Abbreviations 879–885

Absorbance 842

Acetone 246

Acetonitrile 127, 291

Acetonitrile gradient 459

Acetylcholine receptor 533

Acetylcholinesterase 676

α₁-acid glycoprotein 490, 628

Acid precipitation 452

Acidic proteins 273

Acrosin 622

Activated charcoal 267

Activated matrix 707

Activated tentacle chemistry 709, 725

Activated tentacle gels 706

Activated thiol Sepharose 4B 645, 646

Activation by

carbodiimide 719

cyanogen bromide 241, 621–623, 634, 719, 724

divinyl sulfone 621, 624

epichlorohydrine 724

epoxy 719

hydrazide 773

N-hydroxysuccinamide 719, 773

prohormone 655

tresyl 773

Active site 460

Additive(s)

see also Buffers and Detergents, Contents 839–840

additional substances 857–859

antimicrobial agents 860–861

buffer exchange and desalting 859–860

chromatographic modes

biospecific affinity ligands 755, 757

chromatofocusing 395, 404, 406

covalent chromatography 641, 646, 650

dye-ligand affinity chromatography 428–429, 445

hydrophobic interaction chromatography 258

hydroxyapatite chromatography 279, 291, 296

immobilized metal ion affinity chromatography 310, 320, 336–337, 339, 347, 347, 350, 375

integral membrane proteins 529

ion exchange chromatography 6, 8–9, 30, 32–33, 36–39, 48, 62, 77

liquid–liquid partition chromatography 480

reversed phase chromatography 154, 186, 199

scale-up of downstream processing 792–797

size exclusion chromatography 123, 125, 132

elution salts 851

protease inhibitors 854–857

thiol (SH-) reagents 854

Additives 37, 39, 320, 337

Adenovirus attachment protein 537

Adenylate kinase 431

Adenylosuccinate synthetase 431

ADH 486, 489

Adhesion 668

Adhesion efficiency 685–687, 691

Adhesion model of Hammer and Lauffenber 686

Adhesion probability 683–684, 686

Adhesiveness 683

ADP 718

2′, 5′-ADP 306, 433, 718–719, 733, 752–759

2′, 5′-ADP Sepharose 755, 757

Adrenal medulla 410

Adsorption effects 273

Affi-Gel 15, 622, 626, 629, 673

Affi-Gel Heparin 745

Affi-Prep 775

Affinity chromatography 541, 594, 600, 603, 706

Affinity constant 628

Affinity displacing ligands 453

Affinity interactions 465

Affinity matrix 454

Affinity partitioning 437

Agarose 313, 672–673

Agglutinin(s)

see also Lectin(s)

Dictyostelium discoideum 620

Erythrina cristagalli 633–634

Helix pomatia 629, 673–674, 679, 682

Lens culinaris 629

Ricinus communis 620, 629, 633–634

soybean lectin 684

Viscum album 624–625

wheat germ 628

Aggregates 91, 658

Aggregation 291, 334

AH-Sepharose 626

AIA 866

Air bubbles 281

Air degassing unit 94

Air-free conditions 282

Akaline phosphatase 828

Äkta Explorer 791, 810

Äkta 777

D-Alanine carboxypeptidase 532

Albumin 364, 432, 487, 490

Alcohol dehydrogenase 113, 424, 433, 484, 486, 489–490, 492, 500, 719, 753, 759

Aldehyde tentacle gel 714

Aldolase 456–457

Aldolase B 443–444, 454, 466

Alkaline phosphatase 372, 433, 441, 827

Alkyl chain length 532

Alkylating reagents 641

Alkylsulfonates 182

Allyl-2-hydroxy-3-(N,N-dicarboxymethyl)aminopropyl ether 335

Alpha-chymotrypsin 252

Alpha-chymotrypsinogen 252

Alpha-factor receptor (ste2p) (Saccharomyces cerevisiae) 537

Alting-out effect 242

Amberlite XAD-2 530

D-Amino acid oxidase 430

p-Aminophenyl glycosides 621, 626

Amino tentacle gel 715

8-Aminooctyl-Sepharose 4B 293

8-Aminooctyl-Sepharose 352

Aminosuccinic acid 335

Ammonium bicarbonate 183

Ammonium chloride 322

Ammonium hydrogen carbonate 280

Ammonium sulfate

as buffer for hydrophobic interaction chromatography 241–251, 256–267, 572, 576, 850

as other buffer 322, 334, 336, 405, 428, 520, 533, 570–571, 602, 620, 651, 656, 658, 739–741, 757, 799, 850, 851, 860, 862

Ammonium sulfate precipitation 405, 602

AMP 718

5′-AMP 306, 417, 718–720, 723, 733, 752–759, 851

5′-AMP Sepharose 757

Amphipatic 528

Ampholytes 8

Ampholyte displacement chromatography 408

Amphoteric 386

Amphoteric molecules 387

Amphoteric substances 849

Amyloid P component 620

Amylose Resin 592

Analytical Chemistry 151

Analytical Instrument Association 866

Angiotensin derivative 308

Angiotensin II 193

Angiotensinogen 193

Anhydrotrypsin Sepharose 572

Animal lectins 633

Anion exchange chromatography

biospecific affinity ligands 742, 750, 761

buffers and additives 841, 843, 845–849, 852, 858

chromatofocusing 387, 406

displacement chromatography 506, 513, 515, 519–522

DNA-binding proteins 596–597, 603

hydroxtpatite principles 272–273, 297

immobilized artificial membrane chromatography 459, 461, 463

immobilized calmodulin748

immobilized metal ion affinity chromatography 318, 323, 348–350, 355, 361, 363

ion exchange chromatography 7, 17–18, 21–23, 26, 31–33, 36, 38, 40–41, 45, 49, 52, 54–56, 59, 66, 68, 76, 79–81, 540

scale-up of downstream processing 772, 776, 786, 791–793, 798–799, 814, 816

Anion exchange 515, 577, 787, 841, 843

Anion exchanger 18, 79, 273, 461

Anionic 845

Anthraquinone dyes 428

Anti-albumin 487

Antibodies

see also Glycoprotein(s)

chromatographic modes

cell affinity chromatography 668, 671–673, 678, 682, 687–689, 693–696

chromatography on porous matrices 823

chromatography using Strep-tag affinity peptides 826–828

coupling reactions 706, 715

hydrophobic interaction chromatography 237–238, 257, 259, 263

hydroxyapatite chromatography 272, 292

immobilization of nucleotides 718, 721

immobilized metal ion affinity chromatography 310, 348, 364, 366–368, 370

integral membrane proteins 534, 539, 544, 547, 553, 555

ion exchange chromatography 9, 23, 52, 71–74, 76–78

lectins and glycoconjugates 620, 631, 633–634

liquid–liquid partition chromatography 479, 482–484, 494–498, 500–501

recombinant proteins 559, 562–564, 567–568

reversed-phase chromatography 209

scale-up of downstream processing 766, 768, 771, 781

immunoglobulins 73, 75, 119, 237, 306, 308, 335, 364–365, 369, 479, 488, 490, 568, 600–601, 633, 668

IgA 400, 490, 492–493

IgD 208, 220

IgE 676

IgG 21, 74–76, 78, 242, 315–316, 350, 479, 481, 483–484, 489–500, 544, 563, 567–568, 650, 673, 675–678, 688, 823

IgM 119, 238, 490, 492–493, 676

monoclonal antibodies 9, 52, 71–73, 78, 263, 348, 370, 394, 495, 498, 500, 539, 544, 668, 671–672, 682, 766, 823

polyclonal antibodies 238, 500, 534

screening 823

Anti-FLAG M1 affinity gel 566

Antigen(s) 307, 367–369, 495, 497–498, 500–501, 533, 535–537, 541, 544, 549, 559, 637, 658, 694, 823, 827–828, 830, 850

Antigen-binding fragments 544

Anti-IgG 487

Antimicrobial agent(s) 201, 628, 632, 860–861

Antipain hydrochloride 856

Antipapain 625–626

Antiserum 830

Antithrombin III 734–735, 737–738, 743–745, 747

α₁-antitrypsin 489, 490

AP column 802

Ap₆A 510

apoferritin 309

Aprotinin 856

Aqueous polymer systems 474

Aqueous two-phase affinity partitioning 669

Arabinogalactan 620

Arachidonoyl-diacylglycerol kinase 535

Ascites fluid 365

Ascorbate 404, 857

Asialofetuin 627, 634–635

Asialofetuin-sepharose 628

Asialomucin 634

Asialoorosomucoid 627

Aspartase 434

Assembly factor CsgG 827

Asymmetry 130

Atomic absorption spectrometry 323

ATP 718

ATPase 537

ATPase complex 536

ATP-synthase 263

Autoclaving 743

Autoimmune disease 497

Automatic sample injection 283

Automation 691

Autoradiogram 611–613

Autoradiography 605

Avidin 671

Avidin agarose 567

Avidin-activated Biogel P-30 674

Avidin-EDAC Biogel 689

Azlactone 708

Azlactone tentacle gel 715

Azurin 828

B lymphocytes 680

B27 histocompatibility antigen (HLA) 537

Bacillus stearothermophilus 429

Bacillus subtilis 532, 693

Back pressure 51, 112, 275, 279, 288, 296, 389, 788, 798, 806, 811

Bacteria 767

Bacterial protein disulfide isomerase 569

Bacteriophage λ 588

Bacteriorhodopsin 140, 535

Bacteriorhodopsin fragments 538

Bacteriophage P1 607

Band 3 protein from erythrocyte membrane 535

Band broadening or spreading 94, 346, 787, 789, see also Peak broadening

Barley leaves 848

Base matrices 775

Basic proteins 273, 285

Batch experiments 115

Batch processing 874

BBMF 706

BCA assay 452

Bed settling 118

Benzyl alcohol 693

Bestatin 856–857

Betaine 280, 291, 320, 368, 395

Bile acids 66

Bile salts 530

Bi-modal pore media 787

Binding capacity 241, 672, 685, 750, 788

Binding efficiency 809

Bio–Prep SE 105

Bio–Sil SEC 96

Bioaffinity chromatography 541

Bioaffinity ligands 707

BioCAD 777, 791, 810

BioCAD Sprint System 577

Biocompatibility 7

Bio-Gel A 0. 5M 673

Bio-Gel P6 673

Bio-Gel P30 673

Biological activity 841, 850

Biological buffers 845

Biological events 845

Biomolecules 498

BIOSEP–SEC–S400 99

Bio-Sil TSK series 112

Biosilon beads 673

Biospecific affinity gels

see also Contents 731–732

alcohol dehydrogenase 753

application examples 734, 739, 741, 748, 754

calmodulin based affinity chromatography 747

chromatography system set-up 737, 750

growth factors 733, 742

heparin based affinity chromatography 733–747

HIV reverse transcriptase 739

NADPH cytochrome P-450 reductase 754

optimization 737

preparing the gel 748

proteases 750

sample preparation 737, 739, 750, 755

serum proteins 733

steroid receptors 733

troubleshooting 760

Biotin 671

Biotinylated antibody 671, 678

Biotinylated proteins 835

Biotinylating enzyme 568

Bis-tris 397

Bivalent cations 404

Bleeding

coupling reactions 706–707

IMAC 310, 312–313, 318, 323, 335, 337–338, 341, 346, 353, 360, 364–365, 376

Blood platelet membrane 535–536

Blood 679, 695

Blue Dextran 681

Blue Dye 798

Bone marrow 695

Borna disease virus 537

Borosilicate glass 859

Bound proteins 450

Bovine IgG 74

Bovine immunoglobulin 74

Bovine rhodopsin 535

Bovine serum 74

Bovine serum albumin 56, 119, 285–286, 428, 453, 489, 603–606, 624, 671, 678, 857

Bovine viral diarrhea virus 536

BPG 778

Bradykinin 193

Brij 35 530

BSA 487, 489, 671, 678, 798, 800, 805

Buffer(s)

see Contents 839–840

see also Detergent(s)

buffers and additives 840–843, 845–851, 853, 855, 857, 859–861

capacities 31–33, 71, 119–120, 842

choice of 34–36, 38–39, 114–115, 117, 127, 243, 396, 465, 774, 844–846

chromatographic modes

biospecific affinity ligands 735, 739, 748, 750, 754, 756–757, 760–761

cell affinity chromatography 669, 672, 678–680, 689, 691–692

chromatofocusing 387, 392–396, 404–405, 408, 413

covalent chromatography 641, 644, 646, 649, 651, 655, 659, 662, 663

DNA-binding proteins 597, 602

dye-ligand affinity chromatography 428

hydrophobic interaction chromatography 238–239, 243–245, 253, 267

hydroxyapatite chromatography 274, 279–280, 286, 288, 290–291, 293, 296

immobilized artificial membrane chromatography 465

immobilized metal ion affinity chromatography 305, 317–321, 326, 329–330, 332, 336–337, 342–343, 346, 350, 352–356, 358–360, 364, 367, 370, 372, 374

integral membrane proteins 540, 543, 547, 549, 551, 553

ion exchange chromatography 8–9, 13–14, 33, 36, 39–41, 44, 51–52, 58–59, 62, 69, 73–74, 77, 79

lectins and glycoconjugates 621, 628

porous matrices 792–796, 798–799, 809–811, 814

recombinant proteins 561, 564–565, 574, 577, 582–583

reversed-phase chromatography 182, 186, 188–190

scale-up of downstream processing 766, 774, 781–782

size exclusion chromatography 115, 117, 123, 125–127, 132, 140

Strep-tag affinity peptides 830, 834

concentrations 123, 126, 259, 291, 392, 394, 633, 658–659, 674, 678

detection of 48, 245, 267

examples of uses 847–851

exchange 9, 43–44, 93, 95, 287, 859

flow rates 12–13, 131, 253

for lipids and membrane proteins 40, 42–43, 263, 464

ionic strengths 14, 33–35, 305

pH 33, 36, 52, 114, 127, 249, 394, 633, 658–659, 674–678

preparation of 39, 44–46, 117, 186, 245, 279, 319, 325, 329, 396, 680, 735, 748, 754, 799, 846–851

protein determination 245

removal of 287–289, 405, 859

selection 845–846

storage 132, 256, 292

strength 842

theory of 842–845

volatile 35–36, 280

Buffering effect 842

Bulk 275, 280

Bulk gel 342

Bulk materials 8

1,4-Butanediol diglycidylether coupled gel 710

C1* repressor 602

C18 alkyls 517

Ca²⁺ ATPase 535

Cadherin–catenin 573

Calcium chloride 291

Calcium phosphate 272

Calcium-binding proteins 350

Calibration 94

Calibration curve 113, 130

Calmodulin 306, 309, 318–319, 350–352, 562, 565–566, 582, 733, 747–752, 826

Calmodulin agarose 566

Calmodulin based affinity chromatography 747

Calmodulin binding peptide 562, 565–566, 582, 747, 826

Calnexin 463

Calpains 751

c-AMP 718

Candida krusei 305

Candida mycoderma 759

Canine distemper virus 537

Capacities 242, 275, 288, 847

Capillary electrophoresis 78, 94, 376, 513

Capillary electrophoresis apparatus 94

Capillary zonal electrophoresis 814

Caprine arthritis-encephalitis lentivirus 538

Capture (Initial step) 272, 287, 341, 734, 813

Carbamat bond 708

Carbodiimide 626, 707, 719

Carbohydrates 668, 678

Carbohydrate microheterogeneities 217

Carbonyldiimidazol 707, 708

Carboxylates 273

Carboxymethyl 7–8, 22, 26–27, 79, 304, 306, 311, 316, 335, 376, 515, 520–521, 524, 596, 772–793

Carboxymethylated aspartic acid 335

Carboxymethyldextran 8, 515

Carboxymethylpicolylamine 335

Carboxypeptidase G 433

Cardiac membrane proteolipids 538

Carrier 513

Carrier fluids 774

Catecholamine synthesis 410

β-Catenin 573

Cathepsins 655

Cation exchange chromatography 114, 125, 207, 256, 657, 733, 756, 772

buffers and additives 843, 847, 852

displacement chromatography 506, 519, 520–522

DNA-binding proteins 596–597, 600, 603–606

immobilized metal ion affinity chromatography 348, 354, 361

ion exchange chromatography 7–9, 16–17, 21–23, 26–27, 31, 33, 36–38, 47, 53–54, 56, 59, 66, 71, 74–77, 79–81

porous matrices 786–787, 791, 793–794, 820, 822

Cationic 845

CBG 489

Cell affinity chromatography 667–702

see Contents 667–668

application examples 694–696

automation 691

blood 679, 695

CAC kits 678–679

cell preparation 679–680

column testing 681

combinations with other chromatographies 693–694

gels for CAC 672, 674, 687–689

monoclonal antibodies 672, 682

non-specific cell adsorption 670

optional equipment 691

sample loading, washing, and elution 682–683

scaling up and down 690

starting procedures 679

stem cell isolation 695

troubleshooting 696

Cell detachment 684

Cell disruption 561

Cell entrapment 687

Cell function 672, 688

Cell immunoaffinity chromatography 674

Cell preparation 679

Cell removal 684, 687

Cell separation 373

Cell surface probing 373

Cell-CAM 105 535

CELLector Flasks 695

Cellulofine GH-25-c 673

Cellulose 105, 113, 673

Centricon 117

Centrifugation concentration 117

Centroid volume 138

CEPRATE 679, 694–695

CEPRATE Gel 673

Ceramic hydroxyapatite 291

Ceramic materials 275

Ceruloplasmin 490

Cetyltrimethylammoniumbromide 529

cGMP (current Good Manufacturing Practice) 766–767, 782

Channelling 131

Chaotropic 242

Chaotropic agents 396

Chaotropic effect 850

Chaotropic reagents 131

Chaotropic salts 364, 826–827

CHAPS 529—530, 853

Characteristic applications 519

Charge clusters 7

Charged functional groups 387

Charged groups 22

Chelat 650 316

Chelating buffers 291

Chelating Sepharose 566

Chelating Sepharose Fast Flow 314–315, 342, 370

Chelating Sepharose High Performance 324

Chelating Superose 315, 566

Chelation 304

Chelex 100 267

Chicken breast muscle 15

Chicken egg-white cystatin 828

Chimeric IgG antibodies 496

Chitin 620

Chitin beads 565

Chitin binding domain 562

Chlorhexidine 861

Chlorophyll-a/b-binding protein 349, 535

Chloroplast ATPase 61

Chloroplast energy coupling complex 536, 538

Cholate 529, 853

Cholesterol 658

Cholesterol transfer protein 463

Choline acetyl transferase 432

Chromaffin granules 410

Chromaflow 778

Chromatofocusing

see Contents 385–386

application examples 408–413

choice of gel 389, 396

column packing 400

determination of isoelectric points 406–407

ion exchange chromotography 624

marker proteins 400

Mono P 390–391, 409–410

optimal conditions 403–404

pH-gradients 387

principle 387–388

sample preparation 398

scaling up 406

set-up 396–397

test separation 400–402

troubleshooting 258, 413

Chromatography software 863–876

see Contents 863

basic necessities 865–866

commercially-available 868

HPLC software packages 875

interesting features 867

package examples 865

support 875

training courses 875

Chromophore 70

CH-Sepharose 626

Chymase activities 355

Chymosin 430

Chymostatin 625–626, 856

Chymotrypsinogen 244, 247

α-Chymotrypsin 78

α-Chymotrypsinoge 285

α-chymotrypsinogen A 286

Cibacron blue F3G-A 416

CIP

see Cleaning in place

Circular dichroism spectrometer 94

Circular dichroism 140, 142

Citrate 289

Cleaning in place 150, 256, 652, 670, 693, 706, 781

Clostridium perfringens 680

Cloud point 66, 532–533

Cloudy protein solutions 280, 395

CM 7

CMC 64, 123, 529

CMC

see Critical micellar concentration

CMP 718

CM-Sephadex 656–657

CNBr 720, 724

CNBr-activated sepharose 572

CNBr-activated Sepharose 6MB 689

Coagulation factors 306

Coagulation proteins 734

Code system 620

Collagenases 306

Colostrum 726

Column bed height 776

Column bed volumes 780

Column calibration 93

Column conditioning 187

Column cross – sectional area 799

Column diameter 129, 790

Column dimensions 780, 808

Column library 872

Column lifetime 191

Column log-book 811

Column oven 93

Column packing 800

Column size 808

Column switching 319

Column(s)

see also Gel(s)

antimicrobial agents

calibration 89, 91, 93–94, 113, 115, 117–118, 127–128, 130, 133, 143, 188, 239, 246, 326–328, 481, 621, 624, 738, 866

dimension(s) and bed volume(s) 10–11, 15, 25, 42, 46–49, 55–59, 73, 91, 122, 190, 128, 246, 248, 253–256, 260, 262, 264, 282, 285, 292, 320–344, 365–366, 370, 375, 391, 394, 397–400, 403, 406, 409, 413, 439, 474, 479, 486, 508, 517–518, 566, 575, 624–628, 634, 647, 658, 672, 677–678, 680–683, 690–691, 734, 771, 775–781, 790, 798–802, 806–809, 813, 817–821, 831, 833

empty 10, 46–47, 280–281, 342–343, 399, 408, 776–779, 800, 802–803

packing of

cell affinity chromatography 680–681

chromatofocusing 399–400

covalent chromatography 646, 648–649

hydrophobic interaction chromatography 245–246

hydroxyapatite chromatography 276, 280–281

ion exchange chromatography 46–47

reversed-phase chromatography 156, 157, 176, 194

size exclusion chromatography 118

size 11, 117, 128, 244, 251, 259, 261, 290, 336, 404, 445, 459, 486, 517, 776, 781, 808

stabilization 38–41, 95, 622, 754

Column-ovens 155

Columns in tandem 319

Commercially available 437

Compaction 687, 689

Compartmentation 841

Compatibility 864

Competitive ligand 321

Competitive ligand strength 322

Complex protein samples 409

Compression 131, 275

Computer 10, 11, 15, 62, 409, 451, 791, 810

see also Software chromatography software 863–876

Concanavalin 684

Concanavalin A 620

Concanavalin A tentacle affinity matrix 714

Concentration 847

Conductivity detector 513

Conformational changes 500

Conformational structure 237

Connecting a ligand 724–725

Contact time 683

Contain peroxides 851

Continuous bed chromatography 79

Convective mass transfer 789

Convective mass transport 787

Corona virus 533

Corticosteroid-binding globulin 490

Cost(s)

biospecific affinity ligands 742

buffers and additives 854

cell affinity chromatography 668–670, 687, 689, 693

chromatofocusing 408

chromatography on porous matrices 788, 814

chromatography software 867

coupling reactions 706

covalent chromatography 650

dye-ligand affinity chromatography 440

hydrophobic interaction chromatography 238, 251

hydroxyapatite chromatography 275, 290

immobilized metal ion affinity chromatography 310, 314, 342

integral membrane proteins 529, 533–534

ion exchange chromatography 24, 55

recombinant proteins 564, 568

reversed-phase chromatography 194

scale-up of downstream processing 771, 774, 782

software systems

size exclusion chromatography 116, 124, 133, 140, 143

Coulter counters 671

Coupled buffer 289

Coupling reactions

see Contents 705

bleeding 706–707

coupling reactions 706–709, 712–715

spacer technologies 709

synthesis examples 712–715

tentacle gels 711–712, 714–715

Coupling to grafted polymer chains 709

Coupling to spacers 709

Covalent chromatography

see Contents 639

additives

application examples 694–696

direct and analytical modes 646, 648

DTT 654

excess active groups 648

gels commercially available 645

intramolecular disulfide bonds 641

2-mercaptoethanol 641

optimization 651–652

scale-up 652

Triton X-100 652

troubleshooting 662–663

Creatine kinase 433

Critical micellar concentration 52, 63–65, 115–116, 123, 529–532, 541, 548, 852

Cross-linked agarose 95

Crosslinked polyacrylamide 313

Crosslinked polysaccharides 94

Crosslinked polystyrene 95

Crotalus atrox 458, 460–461, 466

Crude egg white 409

Crude extracts 150, 237, 260, 347, 442, 630, 857

Crystallization attempts 141

Crystallization 93, 620, 812

CTAB 65, 852–853

CTP 718

Cuatrecasas 724

Current Contents 151

C_xE_y series 531

Cyanobacteria 814

Cyanobacterial phytochrome 828

Cyanobacterium 295, 349

Cyanogen bromide 239, 241, 572, 595, 597, 600, 621–623, 626–627, 634, 640, 645–646, 659, 672–677, 679, 689, 696, 707–708, 719–721, 724, 744, 747, 748–749, 757, 759, 772

L-Cysteine agarose 645

Cyt b ₆ /f and LCHII 537

Cytochrome b₅ 7, 294, 462–463, 563, 567–568

Cytochrome b-561 410

Cytochrome b ₅₆₂ 828

Cytochrome b ₆ f 294

Cytochrome b ₆ f complex (synechocystis PCC 6803) 535

Cytochrome c 78, 193, 244, 247, 249, 252, 285, 286, 392, 400–401

Cytochrome(s) c

chromatofocusing 392, 400–401, 411, 413

chromatography using Strep-tag

affinity peptides 827–828

hydrophobic interaction

chromatography 244, 246–247, 249–252

hydroxyapatite chromatography 285–286

immobilized artificial membrane chromatography 462

immobilized metal ion affinity chromatography 304–306, 309, 319, 324–326, 330, 332–333, 335, 339, 376

integral membrane proteins 535–538

ion exchange chromatography 67, 78

recombinant proteins 558, 583

size exclusion chromatography 113, 119

Cytochrome c oxidase 535, 558, 827, 833

Cytochrome c oxidase (Bacillus PS3) 536

Cytochrome c oxidase subunits 536, 538

Cytochrome c reductase (potato

mitochondria) (Neurospora crassa) 537

Cytochrome c reduction 462

Cytochrome(s) P-450

biospecific affinity ligands 754, 756, 759

hydroxyapatite chromatography 278, 287–289, 293–294

immobilized artificial membrane chromatography 451, 453, 461–463, 466

immobilized metal ion affinity chromatography 323, 352–355

integral membrane proteins 536, 538, 547–548

ion exchange chromatography 13, 23, 33, 36, 41–43, 48–51, 54–55, 57, 65–67

Cytochrome P-450 fragments 538

Cytochromes (E. coli) 535

Cytolomegalovirus 498

Cytolytic granules 355

Cytomegalovirus 537

Cytosolic proteins 237

Data acquisition 873

Data buffer buffered interface 865

Data-bases for file organization 873

DDE 866

DDT 605

De novo synthesis 138

DE52 cellulose 569, 572

Dead space 131

DEAE 7–8, 18–20, 22, 25–26, 29–30, 40–41, 45, 47, 50–52, 56, 60, 67–68, 223, 258, 308, 350, 361, 370–371, 418–419, 421, 423, 441–442, 444, 463, 481, 511, 519–521, 596, 598–599, 631–632, 646, 649, 654, 656–657, 662, 759, 772, 792–793

DEAE-cellulose 632

DEAE-dextran 8

DEAE-Sephacel 258

Degassing 93, 117, 279, 392, 409

Degradation products 272

Degree of dye substitution 436

Dehydrofolate reductase 661

Dehydrogenases 718

Delipidation 336

Denaturation 291

Denaturing agents 314

Density gradient centrifugation 680

Deoxycholate 65–66, 121, 140–141, 452, 529–530, 534, 537, 629, 633, 853

Depth filter 685

Depth filtration 685

Depyrogenated 692

Depyrogenating 693

Desalting 90, 95, 104–105, 150–151, 183, 217, 257, 323, 336, 338, 340, 348, 438, 512, 589, 597, 604–605, 646, 648, 650, 656, 660, 662, 742, 760, 846, 850, 857–860

Desalting column 340

Desalting in reversed-phase 217

Desthiobiotin 831

Detection 682, 791, 804, 865

Detergent(s)

additives 291

chromatographic modes

affinity chromatography 541–545, 683–689, 751, 759

hydrophobic interaction

chromatography 237–264

hydroxyapatite chromatography 273–289

immobilized artificial membrane chromatography 452–465

immobilized metal ion affinity chromatography 314–344

ion exchange chromatography 9–68, 540–541

reversed-phase chromatography 217–223, 545–547

size exclusion chromatography 92–141, 537–540

cloud point 532–533

compatibility with eluents 66, 652, 804, 852–854

compatibility with gels 67, 249, 320, 337–338, 452–454, 652, 683, 689

concentrations 123–124, 537, 683, 751, 817, 834, 852–854

exchange 117, 140

flow rates 131, 804, 814, 816, 834

general 9, 23, 92, 115, 237–238, 273, 280, 314, 364, 457, 462, 528, 841, 852

interference 454, 529, 561, 622, 629, 751, 814, 816, 852–854

lectins 622–634

membrane proteins 61–68, 115–117, 120–121, 123–124, 281, 528–547, 622, 751, 814, 816, 852–854

microsomes 43, 48, 759

molecular mass 118, 134, 136, 141, 529–530, 532

porous matrices 804–834

properties 61–64, 278, 529–534, 852–854

protein structure or function 65–66, 120

recombinant proteins 561–579

reference reviews 68, 529

removal of detergent

amberlite incubation 530

comparison of gel types 289

by dialysis 529

efficiency 141

eluent properties 287–288

examples 68, 287–288

by hydrophobic interaction chromatography for non-ionics 237

by hydroxyapatite chromatography for non-ionics from membrane proteins 273

important detergent properties 532

by ion exchange chromatography 68, 141

matrix type and pH 58

reference reviews 68

regeneration with ethanol 58

prior to reversed-phase chromatography 217, 223

by size exclusion chromatography 141, 529

wavelengths 67, 339

resolution by 66–67

selection of 33–34, 37–39, 41, 61–64, 115–116, 465, 529, 652

solubility efficiency 64–65, 123–124, 131, 249, 264, 337–338, 465, 561, 579, 622, 629, 652, 683, 751, 759, 814, 816, 827, 834, 852–854

temperature 51, 66, 118, 344, 561, 827, 852–854

wavelength range 67, 116, 287, 339–340, 533, 852–854

Detergent step gradient 460

Detergent-solubilized DMPE 453

Dextran 118, 471, 620, 672–673, 706

Dextran 40 480

Dextran 500 480

Dextransulfate 8

Diabetes mellitus 70

Diafiltration 767

Dialysis 117, 529, 860

Diamid bond 708

Diaminobiotin 565, 827

Diazepam 193

Dichloromethane 132

Dichlorotriazine dyes 436

Dictyostelium discoideum 620

Diethanolamine 402

Diethylaminoethyl 7

Differential refractometer 137

Diffusion paths 787

Diffusion pores 787

Diffusion-controlled process 93

Digitonin 852–853

Dihydrodiol dehydrogenase 660

Dihydrofolate reductase 308, 358–360, 362, 431, 661, 663

Dihydrogenphosphate 279

Dimerization 118

Dimethyl sulfoxide 190, 321, 859

Dinucleotide binding fold 416

Diol tentacle gel 712

Diol 95

Dipeptidyl aminopeptidase 535

Diphenhydramine 193

Dipicolylamine 335, 363

Diptheria toxin 431

2, 2′-Dipyridyl-disulfide 642

Direct coupling 709

Directory of manufacturers and suppliers 887–894

Displacement chromatography

see Contents 505

application examples 522

characteristic applications 519

column size 517

elution modes in 506

gels in 508

ion exchange chromatography 79

location in a purification scheme 508

MALDI-MS 513

principle 506, 508

system set-up 512

troubleshooting 522

Displacement train 509

Displacer molecules 8

Displacing salts 38

Distribution coefficient 91

Disulfide bonds 141, 571, 641

Disulfide bridge 625

Dithiothreitol 37, 39–40, 43–45, 243, 261, 289, 291, 294, 321, 334, 407, 409, 431–432, 435, 545–546, 576, 576, 602, 604, 607–609, 641–642, 648–650, 652, 654–660, 663, 754–755, 841, 854

Divinyl sulfone 621–622, 624

Divinyl sulfone-activated Sepharose 634

DMSO 190, 265, 859

DNA 307

DNA polymerases 739

DNA-binding proteins

see Contents 587

ammonium sulfate precipitation 602

application examples 607–614

autoradiography 605, 611–613

commercially-available gels 598, 600

desalting 604

DNA affinity chromatography 595

DNA mobility shift assay 609

DNA-binding proteins 587–595, 607–609, 611–613

DNase I footprint 611

Heparin Sepharose 597, 589, 602–603, 605

Klenow fragment 609, 611–613

operator 588

protein-DNA interactions 587

purification tag 590

specific site 606

DNase I footprint 611

DNAse 812

DOC

see Deoxycholate

Dodecyl-β-D-maltoside 296, 530

β-Dodecylmaltoside 814

Dodecyl-maltopyranosid 834

Domain-specific binding 7

Donnan potential 114, 406

Dot polymer 711

Downstream processing 706, 826

DP 711

Drivers 866

Dry powder 281

DTE 40, 321

see Dithioerythritol

DTT 40, 289, 291, 321, 654

see Dithiothreitol

Dual-wavelength scanner 452

Duffy protein 658

Dye affinity chromatography 707

Dye leakage 428

Dye-ligand affinity chromatography

see Contents 415

application examples 441–444

column preparation 438–439

column regeneration and storage 440

commercially-available dye-liganded gels 418–426

coupling of reactive dyes 429

dye leakage 428

dye-ligand affinity chromatography 416, 429–437, 441–444

dye-ligand screening kits 437

operational parameters 429

protein purification examples 430–435

reactive dye chemistry 428

sample preparation 437

screening of dye-ligands 442–443

spacer molecules 417

stacking effects 428

troubleshooting 444–445

Dynabeads 373, 695

Dynamic data exchange 866

DNA-binding proteins 589, 593–595, 597, 608–609

dye-ligand affinity chromatography 432, 434

hydroxyapatite chromatography 294

immobilized metal ion affinity chromatography 360–362, 367, 372

integral membrane proteins 535–538

ion exchange chromatography 40

miscellaneous biospecific affinity ligands 739, 741, 747–748, 754–756

recombinant proteins 559–562, 565, 568–569, 571, 573–575, 580

reversed-phase chromatography 215–216

size exclusion chromatography 136

E1 glycoprotein 533

E-64 856

Eddy diffusion 80

EDTA 38, 291, 322, 404, 605, 654, 856–857

Effectively shadowing 685

Efficiency 112

Efficiency-term 128

Egg white 409

EGTA 322

Electron crystallography 814

Electrophoretic separation 386

Electrostatic effect 406

ELISA 529

Eluent 393–394

Eluotropic mobile phase 453–454

Eluotropic strength 452, 458, 464–465, 546

Eluting agents 321

Elution 827

Elution buffer 394

Elution buffer strength 353

Elution chromatography 506, 511

Elution modes 506

Elution salts 851

Elution volume 91

Emulgen 273, 287

Emulgen 911 43, 287, 293, 462

Emulphogen 530

Endcapped 174, 461

Endopeptidases 855

Endoplasmic reticulum 461

β-Endorphin 747

Enterokinase 563

Entrapment of cells 672

Enzymatic digestion 812

Enzymatic lysis 561

Enzyme immobilization 640

Epichlorohydrine 620, 724

Epichlorohydrin-coupled gel 710

Epidermal growth factor 537, 542

Epidermal growth factor receptor 537

Epoxide 292, 798

Epoxy 719

Epoxy-activated gel 710

Epstein–Barr virus 536–537

Equilibration buffer 465

Equilibrium binding capacity 32

Equilibrium constant 137

Equine infectious anemia virus (EIAV) 535

Erythrina cristagalli agglutinin 633, 634

Erythrocyte ghosts 535

Erythrocyte(s) 373, 431, 434, 535–537, 623, 631, 658, 664, 676, 681–682, 692

Erythrocytes (human) (e.g. glycophorin C) 535–537

Escherichia coli 40, 826

Escherichia coli overproducer 589

Ester linkages 460

^esteriam.PC 451, 462–463, 466

Ethanol 291, 861

Ethanolamine 402, 848

^etherIAM.PC^Cl0/C3 451, 454, 456–460, 466

^etherIAM.PE^Cl0/C3 451, 453, 455, 457

^etherIAM.PS^C10/C3 451, 457

Ethylation footprint 613

Ethylene glycol 249, 291, 321, 628

Ethylene oxide 692

Eucaryotes 767

Eucaryotic cells 373

Exopeptidases 855

Expression systems 559

Expression vector 591–593

Extracellular matrix 733

Extraction 531

F₁F₀-ATPase (bovine heart mitochondria) 536

Fab fragments 76

Fab 495

FACS 695

Factionation by LLPC 492

Factor IX concentrate 369

Factor Xa 563

Fast desalting 105

Fc portion 366

FDH 489

Ferric adsorbent paper 373

Ferric ions 499

Ferritin 309

Ferritin 486

Fetal calf serum 678

Fetuin 628

Fibrinogen 306

Fibroblast growth factor 308

Fibronectin 306, 309, 622

Ficoll sedimentation 695

Field flow fractionation 669

Filter membranes 279, 395

Filter 117

Filtration 186, 216, 767

Final polishing 272

FineLINE 778

Fines 281

First separation 284

FITC 688

FITC-conjugated avidin 671

FLAG 560

FLAG-peptide 562, 564

Flavanone 3 β-hydroxylase 407–408

Flow adapter 670

Flow chambers 684

Flow cytometers 671

Flow rate 20, 37, 57, 78, 123, 128, 157, 162, 164, 166, 170, 175, 177, 179, 181, 193, 248, 255, 257, 276, 328, 331, 338, 364–366, 391, 452, 463, 465, 478, 485, 498, 502, 510–511, 516, 519–520, 736, 788–790, 799, 805, 820, 833, 846

Flow-chart 867, 873

Flower color 294, 408

Flow-through system 685

Fluorescence 122

Fluorescence microscopy 682

Fluorescent activated cell sorters 668, 671

Fluorescent activated cell sorting 669

Fluorophore-conjugated antibody 671

FMOC 819

Focused polypeptides 388

Focusing effect 388

Folded protein 560

Folding 142–143

Formaldehyde 693

Forssman glycolipid 629

Four-helix dimer 138

FPLC (fast protein liquid chromatography)

affinity chromatography 740

buffers and additives 860

chromatofocusing 389, 409–410

chromatography on porous matrices 789, 798

chromatography software 865–868

DNA-binding proteins 595, 597

hydrophobic interaction chromatography 239, 244

hydroxyapatite chromatography 274, 276

immobilized metal ion affinity chromatography 311, 313–314, 321, 324–325, 327–330, 338, 341, 343, 345–346, 352–353, 355, 371

ion exchange chromatography 10–11, 13, 15, 25–26, 44, 55, 58–59, 71–72

recombinant proteins 564, 569, 572

FPLC 274, 389

FPLC system 327, 409

Fraction collection 11, 13–14, 16, 48, 74, 122, 155, 247, 259, 263, 274, 311, 324, 339, 347, 389, 401, 472, 518, 569, 577, 597, 619, 647, 691, 734, 737, 748, 750, 754, 791, 804, 810, 863, 866, 869

Fraction collector 14, 791

Fractogel EMD Azlactone 650 (S) 713

Fractogel EMD Chelat 650 (M) 713

Fractogel EMD Chelat 650 (S) 713

Fractogel EMD Epoxy 650 (S) 713

Fractogel EMD TA 650 (S) 713

Fractogel EMD 105, 316

Fractogel 775

French Press 831

Frontal boundary analysis 137

Frontal chromatography 506

Fucan 632

Fucoidan-Sepharose 630, 632

Fumarate reductase 430

Fusion partners 560

Fusion protein 306–308, 357, 590

Fusion protein cleavage 591–593

Fusion 591–593

Fusogenic protein (rat brain microsomal membranes) 537

F_v fragment 367, 833

G1000PW 109

G300SW_XL 99

GAG 746

Galactosamine 679

Galactose-specific proteins 621

β-Galactosidase 309, 563

Galactoside-binding lectin 624

Galactoside-binding proteins 635

β-Galactoside-binding proteins^a 627, 633

Galactoside-specific lectin 625

Galactosyl-specific antibodies 620

Galaptin 633

Galectin-1 625

Galectins 622, 633

Gas chromatography 868–873

Gastric mucin 621

GC200 105

GDH 489

GDP 718

Gel filtration chromatography

see Size exclusion chromatography

Gel filtration experiments 853

Gel filtration 860

Gel matrices 564

Gel permeation chromatography 91

Gel permeation chromatography

see Size exclusion chromatography

Gel permeation 656

Gel suspension 281

Gel(s)

see also Column(s) and Stationary phase(s)

capacity

cell affinity chromatography 671–673, 685–692, 697

chromatofocusing 391–392, 394, 398, 403–405

chromatography on porous matrices 787, 788, 791–792, 794, 796, 798, 806–813, 819–820

chromatography using Strep-tag affinity peptides 831, 835

coupling reactions 706, 709, 713

covalent chromatography 644–645, 648, 651, 654, 660–662

displacement chromatography 507, 513, 515

DNA-binding proteins 594

dye-ligand affinity chromatography 417–426, 436, 438

hydrophobic interaction chromatography 239, 241–242, 252–254, 256

hydroxyapatite chromatography 272, 275–276, 278–279, 290

immobilized artificial membrane chromatography 464

immobilized metal ion affinity chromatography 312–318, 334, 336, 341–343, 346, 350–353, 263–366, 373, 375

ion exchange chromatography 9–11, 17, 19–20, 24, 26–27, 31–33, 35–36, 42, 54–56, 59, 66, 69, 71, 76–77

lectins and glycoconjugates 621–622, 626, 628–630, 634–635

liquid–liquid partition chromatography 475, 481–482

miscellaneous biospecific affinity ligands 733–734, 738, 743–744, 747, 749–750, 752, 757–758

recombinant proteins 566–568

reversed-phase chromatography 152, 157, 160, 162, 164, 168, 170, 176–177, 179, 188–189

scale-up of downstream processing 774, 776, 781

size exclusion chromatography 92, 122, 127–129

commercially-available

2′, 5′-ADP and 5 ′-AMP Sepharose 757

anion exchange chromatography 18–20

calmodulin Sepharose 749

cation exchange chromatography 21–22

cell affinity chromatography 673

chromatofocusing 390

chromatography on porous matrices 792–797

coupling reactions 713

covalent chromatography 645

DNA-binding proteins 598–601

downstream processing for large scale 772–773, 775

downstream processing or technical scale 778–779

dye-ligand affinity chromatography 418–425

heparin affinity chromatographic media 744–745

hydrophobic interaction chromatography 240

hydroxyapatite chromatography 276–277

immobilized metal ion affinity chromatography 315–316, 335

liquid-liquid partition chromatography 473

recombinant proteins 562–563, 566–567

reversed-phase chromatography 157–173

size exclusion chromatography 96–111

equilibration 118, 121, 326, 329, 332, 439–440, 443, 452, 464–465, 480–481, 508, 540, 569, 597, 650, 680, 750, 755, 801

chromatofocusing 392–394, 397, 400–403, 409–410, 413

hydrophobic interaction chromatography 245–246, 250, 259–260, 262, 264

hydroxyapatite chromatography 273, 277, 279, 282, 284–287, 293–294

ion exchange chromatography 9, 39, 41, 44–49, 69, 71, 73, 75

reversed-phase chromatography 152, 180, 187, 190, 192

regeneration 841, 848

cell affinity chromatography 671, 683, 689, 693

chromatofocusing 402–403, 413

chromatography using Strep-tag affinity peptides 831, 833

dye-ligand affinity chromatography 440

hydrophobic interaction chromatography 238, 256–257

hydroxyapatite chromatography 266–277, 291–292

immobilized artificial membrane chromatography 453

immobilized metal ion affinity chromatography 306, 313–314, 321, 324–325, 330, 332, 334, 345, 346

ion exchange chromatography 9, 29, 49, 58–59, 66, 71

lectins and glycoconjugates 628–629, 652

liquid–liquid partition chromatography 488, 509, 512–513

miscellaneous biospecific affinity ligands 743, 748, 751, 758

reversed-phase chromatography 189–190

scale-up of downstream processing 771, 774, 811

size exclusion chromatography 131–132

stabilization 38–41, 95, 622, 754

storage 59–60, 132–133, 191, 256, 291–292, 390, 403, 440, 451, 488, 577, 628–629, 652, 741, 743, 751, 758, 774, 812, 853–858

washing

cell affinity chromatography 679, 681–684, 690–691, 693, 695

chromatofocusing 402, 409, 411–412

chromatography on porous matrices 800–801, 804, 806, 808, 811–813, 827, 829, 831, 834

coupling reactions 706, 716

covalent chromatography 643–644, 646, 651–652

dye-ligand affinity chromatography 428, 439–440, 444

hydrophobic interaction chromatography 243, 247, 256, 259–260, 263–266

hydroxyapatite chromatography 287–288, 291–293, 304, 311, 321, 323, 326, 329, 331, 333, 338, 341, 344–345, 352, 368–370, 375

immobilized artificial membrane chromatography 453–454, 456, 474

integral membrane proteins 544

ion exchange chromatography 13, 44, 4f3–49, 57–58, 60, 69–70, 73

miscellaneous biospecific affinity ligands 751, 755, 758, 761

recombinant proteins 570–572, 576–577, 579, 621, 626, 628, 634

reversed-phase chromatography 187, 190–191, 197

size exclusion chromatography 112, 135

Gelfiltration 405

Generic expression vectors 833

GFC (Gel filtration chromatography)

see Size exclusion chromatography

GFP 831

^δGIAM.PC^C10/C3 451, 460, 454

Glass 673

Glass beads 672, 679

Glass columns 670

Glass matrices 314

Glossary for ion exchange 79–81

GLP (Good Laboratory Practice) 867, 869–873, 875

Glucocerebrosidase (human placenta) 535

Glucokinase 435

Glucose 6-phosphate dehydrogenase 435, 754

Glucose oxidase 72

Glucose transporter protein 463

Glucose transporter 535

Glucose-DH 486

Glutamate dehydrogenase 719

Glutaraldehyde 622

Glutathione 571

Glutathione agarose 567

Glutathione reductase 719

Glutathione Sepharose 590–591

Glutathione-S-transferase 563, 565–566, 659–660

Glutathione transferase 308

Glycerol backbone 460

Glycerol 404, 858–859

Glycerylpropyl bonded surface 112

Glycinehydroxamate 335

Glycoconjugate 625, 726, 713

see also Lectins and glycoconjugates

Glycohistochemical monitoring 632

Glycol 471

Glycoligand 620–621, 630

Glycolipid 629

Glycophorin 373

Glycoprotein D 544

Glycoprotein(s)

see also Lectins and glycoconjugates

ion exchange 40, 67, 69–70, 72

membrane proteins 533, 535, 541, 544

reversed-phase 202, 217, 220–221

Glycosaminoglycan 746–747

Glycosidase 620, 623, 634

Glycosylation 195

Glycosylation patterns 632

Glycosyltransferase 620, 634

Glycyl tyrosine 130

GMP (Good Manufacturing Practice) 706, 718, 726, 875

GMPWXL 111

Goldstein 620

Gonadotropic hormones 306

Good laboratory practice 867

Good-buffer 845, 847

Gossypium hirsutum 463

GPC (Gel permeation chromatography)

see Size exclusion chromatography

Gradient anomalies 737

Gradient conditions 465

Gradient detergent elution 453

Gradient elution 80

Gradient mixer 13

Gradient mobile 452

Gradient salt elution 453

Grafted-tentacle gel 711

Granulocytes 672

Granzymes 354

Grapevine leaf-roll associated virus 859

Gravity flow 670

Green fluorescent protein 831

Growth factors 306, 733, 742

Guanidine hydrochloride 743, 835, 859

Guanidine 131, 141, 292, 314

Guanidinium chloride 38

Guanidinium hydrochloride 127

Guard columns 452

Guideline for binding and elution 286

HABA 831

Haemacytometer 671

Haemagglutination assays 623, 630

Haemolysin 658

Halobacterium halobium 140, 535

Halogen containing compounds 114

Halophilic bacteria 9

Handbook templates 872

Hanks balanced salt solution 678

Haptens 498

Hard metal ions 350

Hardware 864

HA-Ultrogel 293

HBGFs 742

Hecameg 530

Helix pomatia agglutinin 629, 673–674, 679, 682

Helix pomatia-Sepharose 689

Heme 70

Heme groups 791

Hemiacetal hydroxyl group 621

Hemoglobin 70

Henderson and Hasselbalch 842

Heparin 798

Heparin based affinity chromatography 733–747

Heparin-binding growth factor 742

Heparin-binding lectin 623, 625

Heparin HyperD M 744

Heparin Sepharose 589, 597, 602–603, 605, 623, 744

Hepatic triglyceride lipase 734

Hepatitis B 537

Heptafluorobutyric acid 182

Herpes simplex virus 535–538, 544

Heterobifunctional crosslinking reagents 641

Hexokinase 430

HFBA 182

Hibitane digluconate 693

HIC

see Hydrophobic interaction chromatography

(His)₆-tag 592

His₆; His₁₀ 562

His-bind 566

His-patches 564

His-tagged proteins 307, 357, 564

Histamine 672

Histidine 392

Histidine residues 309

Histones 202, 392

HiTrap Chelating 315

HiTrap columns 313

HiTrap Desalting 104

HiTrap Heparin 744

HIV reverse transcriptase 739

HLB values 64

HLB 529

see Hydrophilic–lipophilic balance

HMG-box 575

Hofmeister series 248, 850

Hollow fiber membranes 691

Horse 304

Hotlines 875

HpA-Biogel P-30 687

HpA Sepharose System 689

HpA-Sepharose 6MB 681

HPLC software packages 875

HPLC

affinity chromatography 537, 541–545, 547

buffers and additives 860

chromatofocusing 389

chromatography on porous matrices 789–791, 799–804, 806, 810, 813–814, 818

chromatography software 864–865, 868–872, 874–875

displacement chromatography 507, 511–514, 518, 528

dye-ligand affinity chromatography 428

hydrophobic interaction chromatography 238–239, 244, 247, 259

hydroxyapatite chromatography 274, 276, 278, 534

immobilized artificial membrane chromatography 451, 463–464, 466

immobilized metal ion affinity chromatography 311, 313–314, 321, 324–328, 341–344, 352, 365, 372

integral membrane proteins 528, 534–549

ion exchange chromatography 11, 12, 15, 25–26, 39, 44, 51–52, 55, 58–59, 70, 74, 76, 79–80, 536, 540–541, 547

liquid–liquid partition chromatography 472, 481, 489–490

recombinant proteins 564, 569

reversed-phase chromatography 150–151, 153–155, 178–179, 185–187, 189, 191–192, 194–195, 538, 541–545–547

scale-up of downstream processing 766, 774, 777

size exclusion chromatography 91–95, 115–118, 120, 122–127, 129–130, 132–134, 136–137, 140–143, 535, 537–539, 547

HPLC-grade water 186

HPSEC 91

HR Column 803

HSA 489, 678

Human lung leukotriene C₄ synthase 537

Humanized murine IgG₁ 365

Hybrid ions 291

Hybridoma technology 766

Hydrazine 798

Hydrodynamic diameters 90

Hydrodynamic instability 849

Hydrodynamic viscosity radius 27

Hydrodynamic volume 91

Hydrogenphosphate 843

Hydrophathy plots 252

Hydrophilic heads 528

Hydrophilic vinyl–polymer 108

Hydrophilic–lipophilic balance number 531

Hydrophilicity 531

Hydrophilic–lipophilic balance 63–64, 529, 531–532, 548–549

Hydrophobic adsorption chromatography 236

Hydrophobic amino acid 236

Hydrophobic domains 852

Hydrophobic interaction chromatography 450

see Contents 235–236

ammonium sulfate 261, 267

application examples 258–266

appropriate buffers and salts 241–244

buffers 243

cleaning in place (CIP) 256

column and gel maintenance 256

column calibration 246

detergents 249

ethylene glycol 249

gels, syntheses and capacities 239–241

hydroxyapatite principles 273

immodulised calmodulin 748

optimization of running conditions 248–250

packing the column 245–246

preparation of samples 244

recombinant proteins 237, 571

salting-out effect 242

sample properties 256

system set-up 244

troubleshooting 257, 266–267

Hydrophobic interactions 114, 628

Hydrophobic proteins 807

Hydrophobic salting-out chromatography 236

Hydrophobic tail 528

Hydrophobicity 62, 149, 532, 850

Hydropore 461

Hydropore anion-exchange column 459

Hydroxamic acids 335

Hydroxyapatite 594, 814

Hydroxyapatite chromatography

see Contents 271–272

application examples 292–297

back pressures 275, 279, 288, 296

capacities 288

column packing 280–281

commercially-available gels and columns 275

cytochrome P-450 287

phosphate 279–280

removal of detergent 278, 287–288

sample preparation 280

set-up 279

test mixture 284

troubleshooting 296–297

volatile buffers 280

4-Hydroxybenzoic acid 193

Hydroxylapatite 271

Hydroxylated polyether copolymer 127

Hydroxylation 292

12′-Hydroxysteroid dehydrogenase 434

Hygiene 343

HyperD 315, 787, 791, 807

Hyper D F 775

HyperD gels 315, 335, 744, 787, 791–793, 795, 797–798, 807–809

HyperD media 335

Hyperglycemia 70

IAMC

see Immobilized artificial membrane chromatography

IDA

see Iminodiacetic acid

EEC

see Ion exchange chromatography

IEF

see Isoelectric focusing

IgA 493

IgG 482, 484, 487, 493, 496, 823

IgG₁ 366

IgG Sepharose 567

IgM 493

IMAC

see Immobilized metal ion affinity chromatography

IMAP 373

IMEC 334

Imidazole gradient 322

Imidazole 289, 321, 332, 351, 392, 397

Iminodiacetic acid 34, 304–324, 334–335, 338, 344, 351, 353, 355, 358–360, 365, 367–370, 372–376, 394, 397, 520, 707, 711, 713, 715, 719

Iminodiacetic acid 304, 311

Iminodiacetic acid 707

Iminodimethyldiacetic acid 335

Iminodipropionic acid 335

Immbolized phoshpolipids 457

Immobilized antibodies 706

Immobilization of nucleotides

see Contents 717

connecting a ligand 724–725

linking 719

new applications for nucleotide gels 726–727

purification using nucleotides 719

solid phase syntheses using Fractogel EMD Epoxy 726

toxicity 720

Immobilization of

2′, 5′-ADP 759

5′-AMP 759

antibodies 718

calmodulin 748

dehydrofolate reductase 661

disulfide 643

heparin 747

metal ions in IMAC 317–319

nucleotide sugars 720

phosphate group 720

Protein A 718

Protein G 718

proteins on an azlactone tentacle gel 715

support materials 721

Immobilized artificial membrane

chromatography

see Contents 449

cytochrome P-450 466

detergent step gradient 460

endcapped 461

equipment set-up 464

integral membrane protein 464

large-scale purification 463

preparative-sized column 463

protein content measurement 452–453

purification examples 453–464

sample preparation 451

Immobilized iminodiacetic acid tentacle gel 715

Immobilized lipid 464

Immobilized metal ion affinity chromatography

see Contents 301–302

antibodies 364

application examples 348–371

automation 345–346

bleeding or leakage 312, 318, 323, 337–338, 342, 364

buffer solutions for a pH gradient 329

commercially available gels 314–317

cytochrome P-450 isozymes 352–354

cytochromes c 302, 304–306, 309, 319, 324–325, 327

detergents 337, 364

eluting agents 321–323

enzyme activity, binding, inhibition, stability and topography 308–309

Fractogel EMD 316

glycoproteins 308

HiTrap columns 313

IDA 304, 311, 360

immunoglobulins 306, 308, 364

large-scale preparation of factor IX concentrate 369–371

membrane proteins 306, 541

metal ions 317–319

metalloproteins 306, 318, 372

monoclonal antibodies 370

new and unusual gels 335

NTA 304, 311, 359–360

optimizing 334–341

photosystems I and II 306, 348

pre-bleeding 323, 338, 353

principles 303–350

recombinant and fusion proteins 307–308, 324, 335, 337, 357–364, 564

scale-up 342

scaling down 342

SMART system 325

TED 304, 311

tentacles 313

test run using a syringe 330–333

test run with an FPLC system 327–330

test run with an HPLC system 325–327

trace enrichment 346

troubleshooting 374, 376

Immune mediator release 633

Immunoaffinity chromatography 678

Immunobac 544

Immunoglobulin G 633

Immunoglobulins 306, 308, 364

see also Antibodies

Immunomagnetic bead depletion 669

Immunomagnetically labeled cells 691

Immunoprotein affinity gels 364

IMPACT 560, 575

Inclusion bodies 579, 766

Incubation time 683

INdEX 779

Induction 591–593

Industrial downstream processes 718

Industrial preparative separation 875

Influenza virus 535, 538

Initial breakthrough concentration 685

Initial capture 272

Injection valve 93

Inlet and outlet ball 94

Inner mitochondrial membrane 461

Inorganic matrix 272

Insect transferrin 363

Insulin B-chain 193

Insulin cleaving proteinase 264

Integral membrane protein 658, 841

see Contents 527

application examples 537–547

bioaffinity chromatography 534, 537

biological activity 534

cloud point 532

critical micellar concentration (CMC) 529–530

cytochrome P-450 536, 538

detergents for membrane proteins 464, 529–530

ELISA 529

hydrophobicity 532

ion exchange HPLC 534, 536

micellar molecular weight 530, 532

monoclonal antibodies 544

phospholipid 528–529

price(s) 533

reversed-phase HPLC 534, 538

size exclusion HPLC 534, 535

troubleshooting 548

UV-transparency 533

Integration 874

α₁β₁-Integrin 535

Interfacial enzymes 453

Interferon 306, 308, 432, 747

Intermolecular dioxygenases 294, 408

Intestine 441, 463

Intestinal peptide transporter 536

Intramolecular disulfide bonds 641

Ion exchange buffers 33

Ion exchange chromatography 594, 598, 624–625

see Contents 3–4

see also Anion exchange chromatography

see also Cation exchange chromatography

β-lactoglobulins 51

additives 37, 39

anion exchangers 18

antibodies 76

application examples 70–78

buffers 34

capacity 31–32

cation exchangers 21

chromatography on zirconia 78–79

cytochrome P-450 41

detection 48

detergent removal 68

detergents 38, 63, 67

eluting agents 35–37

equipment 10–15

Fab fragments 76

gel itself 16–24, 54–55

glycoproteins 69–70

HPLC 80

hydrophilic–lipophilic balance 64

hydrophobicity 62

isozymes 41, 72

liver microsomes 41

optimization 49–52

packing the column 46–47

particle size and porosity 24–28

porosity 6

purification of immunoglobulin G₁ 74–76

sample preparation 42–44

scaling up and down 56–57

SNAP 33, 56, 74, 80

strong ion exchanger 80

system set-up 41–42

tentacle ion exchangers 7, 55

troubleshooting 77–78

UV/VIS-monitors 13

weak ion exchanger 81

xenobiotics 42

Ion exchange 273

Ion exchange HPLC 540

Ion exchanger 387, 596

Ion pair reagent 149

Ionic capacity 80

Ionic effects 114

Ionic interactions 114

Ionic strength 280, 387, 395

Ionotope 7, 80

Ion-pair model 153

IPTG 569, 573, 575, 597, 604

Irreversible binding 150

Isocitrate 404

Isocitrate dehydrogenase 719

Isocratic formation 387

Isocratic pump 94

Isoelectric focusing 70, 73–74, 76, 207, 212, 222, 386–387, 489, 492, 495, 499, 675

Isoelectric point 114–115, 119–120, 181, 249, 257, 386, 479, 490, 499, 565, 630, 742, 752, 780, 792, 841, 847, 849

Immobilized metal ion affinity chromatography 305, 310, 319, 361, 386–389, 392, 396, 400, 404, 406–410, 412

Ion exchange chromatography 7, 18–19, 22, 32, 47, 71, 73, 78, 80

Isoelectric variants 625

Isoenzymes 272, 292, 404, 462

Isoforms 292, 404, 624

Isoforms of monoclonal antibodies 71

Isopropanol 116, 133

Isothiocyanate 626

Isourea 634

Isourea bond 708

Isozymes 41, 72

Kinases 718, 752

Kininogen 306

Klenow fragment 609, 611–613

Kyte–Doolittle plot 62

Lactate dehydrogenase 430, 434, 492, 499, 719, 753

Lactoferrin 306, 318, 431

β-Lactoglobulins 51

Lactose-Sepharose 624

LALLS 94, 137, 238

LALLS photometer 137

Lambda repressor 588

Large proteins 218

Large scale 57, 72, 156, 306, 360, 369–370, 417, 463, 559–560, 565, 657, 734, 766–767, 769–772, 774, 781, 782, 788–789, 798, 813, 815

see also Scaling up

Large zone 138

Large zone size exclusion 137

Large-scale preparation 369

Large-scale purification 463

β₁-Latency-associated peptide (β₁-LAP) 435

LC-GC-online 151

LDH 489

Leaching 312

Leakage 312, 323, 743

LEC 303

Lectin activity 629

Lectin blotting 622

Lectin(s)

see also Agglutinin(s)

see also Glycoconjugate(s)

see also Cell affinity chromatography

agarose gel 680, 687

animal 627

calcium binding 307

carbohydrate combination 678

coupling reactions 706

galactoside-binding 624

general 620–622

heparin-binding 623

horse gram 317

IMAC

mistletoe 627

plant 627

serial lectin affinity 620, 630

soybean 684, 688

tissue 630

Urtica dioica 623

wheat germ 628, 673

Lectin-bound cells 678

Lectin-containing gels 622

Lectins and glycoconjugates

see Contents 619

agglutinins 620

application examples 632–634

combination with other chromatographies 630

glycohistochemical monitoring 632

lectin purification modification 626–628

lymphocyte migration 632

NMR spectroscopy 620

standard purification scheme 623, 625

sugar receptor purification 632

troubleshooting 634–635

LEF-1 575

Leishmania membrane protein 536

Lens culinaris agglutinin 629

Lentil lectin 401

Leukaemia cells 355

Leukocytes 679

Leupeptin 625–626, 856

Lewis acid 79, 304

Lewis base 79, 304, 312, 347

LHRH 308, 358

LiChrospher-Diol 472

LiChrospher Diol 4000 478, 483, 489

Ligand blotting 626

Ligand densities 237, 709

Ligand exchange chromatography 303

Ligand immobilization 672

Ligand(s)

cell affinity chromatography 668–669, 671–672, 674–676, 678–679, 683–684, 686–689, 692–695

chromatography using Strep-tag affinity peptides 823, 827, 831

coupling reactions 706–716

covalent chromatography 640, 643–646, 648–650, 652–653, 659–660

DNA-binding proteins 594, 600–601

dye-ligand affinity chromatography 416–445

hydrophobic interaction chromatography 236–241, 248, 250–252, 266

immobilization of nucleotides 717–720, 719–726

immobilized artificial membrane chromatography 453–454, 466

immobilized metal ion affinity chromatography 303–306, 311–312, 317–318, 321–323, 349–350, 353, 364–365, 376

integral membrane proteins 536, 541–548

ion exchange chromatography 8–9, 22, 41

lectins and glycoconjugates 620–623, 625–630, 632, 634–635

liquid–liquid partition chromatography 471, 486, 490, 494, 498–500

miscellaneous biospecific affinity ligands 732–734, 742–754, 757–761

scale-up of downstream processing 771–772, 781

Light chain 366

Light-harvesting complex proteins 816

LIMS 865

Linear flow rate 80, 776

Linear gradient 809

Linear velocity 799

Linking 719

LiParGel 650 472, 478, 483, 489

Lipid environment 287

Lipids 40, 140, 323, 528, 811

Lipoproteins 733

Lipoprotein Upase 734

Liquid–liquid partition chromatography

see Contents 469–470

antibody difference detection by LLPC 484, 498

application examples 488–508

chromatograms in LLPC 483

column dimensions and scaling up and

down 486

gels 472

IgG 482, 484, 493, 496

LiChrospher Diol 4000 478, 483, 489

monoclonal IgG 495–498

optimizing of running conditions 482

partition properties for LLPC 487

PEG 8000 480

polyclonal IgG 491, 494

sample preparation 479

surface properties 498

surface property comparison by LLPC 497–498

system set-up 475

troubleshooting 501–502

Lithium chloride 852

Lithium salts 851

Lithiumdodecylsulfate 135

Liver microsomes 41, 293, 462

Liver 443

LIVM values 62

LLPC

see Liquid–liquid partition chromatography

Loading capacities 809

Low-angle laser light-scattering 136

Low angle laser light scattering photometry 238

LPLC (low pressure liquid chromatography)

immobilized metal ion affinity chromatography 324, 330–333

ion exchange chromatography 10–11, 13–14

Lubrol 852–853

Lubrol PX 43, 530

Lymphocytes 356, 533, 672, 675, 679

Lymphocyte migration 632

Lymphokynes 694

Lyophilization 217, 846

Lysozyme 244, 247, 249, 252, 285, 306, 309

l-Cysteine agarose 645

Macromolecules 289

Macroporous gels 709

Macro-Prep 775, 787, 791, 807

Macro-Prep gels 791–798

Magnetic activated cell sorter 691

Magnetically stabilized fluidized bed 692

Magnetized steel wool columns 691

Malate dehydrogenase 492, 753

MALDI-MS 155, 513

Maltose binding protein 562, 565

β-D-Maltoside 288

Mammalian cells 767

Mammalian galectin-1 633

Manduca sexta 363

Mannan 635

Mannan-binding protein 620, 622

Mannitol 115

Mannitol-specific enzyme II of the E. coli 535

Mannose 634

Mannose-6-phosphate receptor 634

Manufacturers 887–894

Marker proteins 400

Mass recovery 241

Mass spectrometer 154

Mass spectrometry 143, 195, 620, 620

Mass spectroscopy 140, 142

Mast cells 676

Matrix 336, 417, 473, 709

Maximizing resolution 288

MDH 489

Measles virus 536–537

Mecuric reductase 431

MedLine 151

Membrane affinity chromatography surfac 454

Membrane antigen (Plasmodium falciparum) 536

Membrane filters 285

Membrane glycoprotein antigen 535

Membrane multiprotein complex 814

Membrane protein complexes 835

Membrane protein solubilization 854

Membrane protein 143, 222, 273, 306, 410, 453, 536–538, 806–807, 811

Membrane 450

Membranes (E. coli) 536

Mercaptoethanol 854

2-Mercaptoethanol

covalent chromatography 641–642, 648–650, 661–663

dye-ligand affinity chromatography 430–431, 433–434, 443–444

hydrophobic interaction chromatography 243

immobilized artificial membrane chromatography 456, 466

immobilized metal ion affinity chromatography 303, 320, 321, 334, 376

ion exchange chromatography 40, 77

lectins and glycoconjugates 621, 623, 626

miscellaneous biospecific affinity ligands 739, 841, 854

recombinant proteins 576

size exclusion chromatography 98–99, 105

Merthiolate^® 627, 737

Metabolic pathways 841

Metal ion 486

Metal ion leakage

see Bleeding

Metal ion transfer 372

Metalloproteases 855

Metalloproteins 306, 318, 372, 827

Metastasis formation 632

Methanol 132, 291

Methylation 614

Methylation footprint 612

Methyl-Sepharose 748

MHC class II-peptide antigen complexes 535

Micellar molecular weight 530, 532

Micelle size 116

Microbial contamination 402, 812

Microbial growth 59, 310

Microbiological growth 194

Microbore columns 154

Micrococcus luteus 532

Microfiltration 767

Microheterogeneity 142

Microsomal membrane proteins (rat liver) 536

Microsomal proteins 462

Microsomes 42

Microviscosity 457

MiniMACS 695

Mistletoe lectin 627

MIT 372

Mixed disulfide bonds 641

Mixed-bed gels 143

Mobile phase system 180

Mobile phase 5, 465

see also Buffer(s)

Mode of branching 630

Model membranes 140

Modifier 114, 457

Modifying agents 320

Moduline acrylic 779

Moduline ss 779

Molecular maquettes 138

Molecular mass determinations 93

Molecular size 90

Molecular weights 90

Moloney murine leukemia virus 538

Mono and dinucleotides 307

Mono P

chromatofocusing 390–391, 394, 397–398, 400, 402, 406–407, 409–410, 412

DNA-binding proteins 599

Mono Q

buffers and additives 848–849

DNA-binding proteins 599

hydroxyapatite chromatography 297

immobilized metal ion affinity chromatography 320, 355

integral membrane proteins 542

ion exchange chromatography 15, 19, 26, 29–30, 36, 40, 47, 49–50, 53, 57–59, 66–68, 72–74

recombinant proteins 577–578

Mono S

displacement chromatography 519

DNA-binding proteins 589, 597, 599, 604–606

ion exchange chromatography 15, 21, 26, 30, 40, 47, 58–59, 67, 71–72

Monoamine transporter 535

Monoclonal antibodies 370, 544, 672, 682, 823

Monoclonal IgG 495, 498

Monoclonal IgG2b 496

Monocytes 672

Monohydrogenphosphate 279

Mononucleotides 307

Monooxygenases 292

Motional properties 457

MPLC (medium pressure liquid chromatography)

see FPLC (fast protein liquid chromatography)

MS

see Mass spectrometry

Mucin 621, 627, 631–632, 634

Multicolumn methods 866

Multimedia-HPLC 875

Multi-receptor attachment 684

Multi-tasking systems 865

Murine IgG₁ 365

Muscarinic acetylcholine receptor 535

Mycoplasma gallisepticum 535–536

Mycoplasma hyopneumoniae 538

Myc-tag 826

Myelin glycoprotein P₀ 538

Myeloma IgG 497

Myoglobin 78, 244, 247, 252, 306, 309, 489, 790

Myosin 432

Myosin light chain kinase 747

N-(4-vinylbenzyl)-imino]diacetic acid 335

N ⁵-methyltetrahydromethanopterin: coenzyme M 535–536

N-acetyl-cysteine 571

N-acetylgalactosamine 620, 629, 681

N-acetylneuraminic acid 632

N-acetylphosphatidylethanolamine synthase 453, 455, 466

N-hydroxysuccinimide 622, 640, 659, 719, 744, 773

n-octyl-β-D-glucopyranoside 116

NAD 601, 757

NAD⁺ 429, 434, 747, 753–754, 718–719

NADH 294, 430, 435, 440, 450, 499, 535, 718, 733, 747, 752–754, 757–758

NADH oxidase 430, 535

NADH⁺ 499

NADH/NAD⁺ 718

NADP 6, 67, 757

NADP⁺ 435, 660–661, 718–719, 753,

NADPH 431, 718, 733, 747, 752–754, 756–759, 851

NADPH cytochrome P-450 reductase 754

NaDPH/NADP⁺ 718

Nanofiltration 767

NAPES 466

Naphthalene dyes 428

Natural killer cells 678, 688

Negative purification 319

Negative selection 679, 688

Negative selection panning 694

Neopentyl 240

Neuraminidase 679–680

Neuronal cells 676–677

Neuropeptides 195

NHS

see N-hydroxysuccinimide

Ni²⁺-IDA 360

Ni²⁺-NTA 360

Ni-NTA 316

Ni-NTA agarose 566

Ni-NTA Silica 316, 566

Ni-NTA superflow 566

Nitric acid 129

Nitroloacetic acid 304, 306–308, 311–313, 316–317, 321, 334, 344, 358–363, 373–374, 592

NMR 142, 471, 573, 588, 814

NMR spectroscopy 620

Non-denaturing 852

Non-fusion recombinant proteins 360

Nonidet P40 530

Non-ionic detergents 273, 404

Non-polymeric focusing 395

Non-selective desorption 457

Non-specific binding 672, 685

Non-specific cell adsorption 670

Non-specific cell entrapment 692

Non-specific ionic interactions 319

Novarose 364

NP-40 410, 529

NTA

see Nitroloacetic acid

Nuclear magnetic resonance

see NMR

Nucleic acid binding proteins 733, 738

Nucleobase-linked nucleoside phosphates 721

Nucleosid diphosphate kinase 434

Nylon wool column 695

Object linked embedding 865

Octyl glucoside 320, 853–854

Octyl-glucopyranosid 834

Octylglucoside 529–531

OLE 865, 875

Olfactory neuronal transcription factor 614

Oligomer 123

Oligomerizations 120

Oligosaccharides 621

OmpA signal sequence 571

OMPs (E. coli) 536

OMPs of Haemophilus influenzae 535

On-column cleavage 576

Oncoprotein bcl-2 537

One-hybrid system 588

On-line dynamic light scattering detector 94

On-line light scattering detector 93, 143

Online low-angle laser light scattering detector 94

On-line viscometer 93

Operating system 865

Operator 588

O-phosphoserine 335

Optical filter 328

Optical flow-through cell 93

Optimization 188, 810

Organic bonded phases 125

Organic compounds 291

Organic modifier 546

Organic polymers 92

Organic solvent elution 453

Organic solvents 40, 127, 131, 280, 403, 457

Organosilane 621

Orotate phosphoribosyltransferase 432

OS/2 866

Osmotic shock 561

Outer membrane proteinase 264

OVA 489

Ovalbumin 50, 252, 635, 798, 800

Ovalbumine 805

Ovomucoid 621, 628

Oxamate 499

Oxirane 708, 724

Oxirane-activated matrices 241

Packing of column(s)

see Column(s), packing of

Packing reservoir 281, 399

Packing state 131

Packings 240

Pancreatic PLA₂ 454, 466

Panning 669, 695

Papain 650, 855

Papain-digestion 77

Particle size 178, 281, 809

Partition coefficient 471

Pass through 282

Pass through peak 465

PBE

see Polybuffer exchanger

PC data use 865

PD10 column 117

Peak broadening

chromatography on porous matrices 787, 789, 809

immobilized metal ion affinity chromatography 338, 346

ion exchange chromatography 26, 28, 46, 49, 80

reversed-phase chromatography 188, 202

size exclusion chromatography 118, 125, 128, 130, 132, 189

Peak capacity 92

Peak labeling 874

Peak splitting 131

Peak volume 112

PEEK 154, 200, 790–791, 801, 803, 824, 864

Pefabloc SC 855–856

Pefabloc^® 739

PEG 8000 480

PEG

see Polyethylene glycol

Penicillium amagasakiense 53, 72

PEPcK

see Phosphoenolpyruvate carboxykinase

PEP-dependent phosphotransferase system> 535

Pepsin 392

Pepstatin 856–857

Peptide hydrolase 653

Peptide mapping 208

Peptide maps 202

Peptides 121, 138

Peracetic acid 693

Perchlorate 183

Perforins 354

Perfusion chromatography 80, 176, 787–788

see also Chromatography on porous matrices and POROS gels

Peripheral basophils 676

Peripheral blood cells 677

Peripheral blood lymphocyte 695

Peripheral membrane protein 293, 454, 528

Periplasmic proteins 561

Peristaltic equipment 789

Peristaltic pumps 670

Peroxidase 489

pH

adjustment of for size exclusion chromatography runs 114, 849

analysis of for protein folding by size exclusion 141–142

changing pH for optimization of hydrophobic interaction chromatography 249, 850

changing pH for optimization of membrane protein separation 810

choice of for 5 ′–AMP and 2′, 5′-AMP runs 757

choice of for affinity chromatography 850–851

choice of for covalent chromatography 651

choice of for immobilized calmodulin 750

descending pH gradients in immobilized metal ion affinity chromatography 327, 329, 338

for carrier support of pH in displacement chromatography 513, 515

for influence of on binding to Mono Q 848

for protein folding and stability with Strep-tag affinity 826

for use in buffers 842–843, 845–847

pH gradients in ion exchange chromatography 52, 66, 71, 73, 76

pH limits in ion exchange chromatography 32, 36

pH of aqueous components for reversed-phase chromatography 180

pH regions in chromatofocusing 356–357, 849

silica-based sorbent choices for reversed-phase chromatography 186

pH-changes 290

pH interval 408

pH monitor 327

Pharmalyte 393–394, 849

Phase separation 853

Phases 452

Phenol red 400

Phenol(resorcinol)-sulfuric acid assay 628

Phenyl sepharose 570

Phenylarsine oxide 568

Phenylglyoxal 622

Phenylmercury salts 861

pH-gradients 387

Phosphatases 306, 752

Phosphate buffered saline 678

Phosphate 115, 127, 279–280, 843

Phosphatidylcholine 451, 453–454

Phosphatidylethanolamine 451, 453–456

Phosphoacetylmuramylpentapeptide translocase 532

Phosphoamino acids 322

1,6-Phosphoaminohexyl spacer 759

Phosphoenolpyruvate carboxykinase 260–264

Phosphofructokinase 416

6-Phosphoglucanate dehydrogenase 754

6-Phosphogluconate dehydratase 433

Phosphoglycerate kinase 434

Phospholipase A₂ 454

Phospholipid 528–529

Phospholipid bilayer 451

Phosphopeptides 154, 318

Phosphoproteins 306, 322

Phosphoramidon 856

Phosphoric acid 843

Phosphorlyated nucleotides 752

Phosphorylation 634

Photo diode array detector 93

Photoautotrophic organisms 294

Photosynthic membrane proteins 806–809

Photosystem 67–68, 121, 123–124, 136–137, 296, 306, 535, 7, 828

PS 1291, 295, 297, 348–350, 814, 816–818, 824

PS II 294–295, 348–350

Photosystem II 294

Phthalocyanine dyes 428

Physiological conditions 842

Phytochrome 431

pI

see Isoelectric point

Pigment 121

Pilot chromatographic studies 464

Pinpoint peptide 563, 568

PIPES 291

Piston seal rinse 94

pK _a 33, 40, 842

pK _a-values 52, 66

PLA₂ 456, 458–459, 461, 466

Placenta 623, 626, 631–633

Plasma 370

Plasma membrane proteins 543

Plasma membrane proteins (human) 537

Plasma membranes (liver)/morris hepatoma 535–537

Plasma protein 511

Plasmacytoma cells 677

Plasminogen 434

Plasminogen activator 306

Plasmodium vivax 658

Platelets 677

Platelet-derived growth factor receptor 535–536, 538

Plugging 811

Pluronic F68 363

PMSF 37, 39, 41, 43, 293, 320, 348, 359, 361, 754, 841, 855–856

Polarity of solution additives 859

Polishing (Final step) 150, 272, 287, 323, 341, 348, 653, 742

Poly(2-hydroxyethyl methacrylate 694

Polyacrylamide 621, 672–673, 706

Polyamine graft copolymer 694

Polybuffer 393–394, 405, 849

Polybuffer exchanger 94, 118, 258–259, 390–393, 397, 399–403, 406, 408, 410, 411, 861

Polychlorosilanes 174

Polyclonal IgG 491, 494

Polyclonal immunoglobulins 365

Polyethylene 471

Polyethylene glycol 39, 677, 860

dye-ligand affinity chromatography 437–439, 442–444

immobilized metal ion affinity chromatography 334, 336, 373, 376

liquid–liquid partition chromatography 471, 474, 476–480, 483–487, 489–497, 499, 501

Polyethylene glycol precipitation 336

Polyethyleneimine 594

Polyethylenimine-modified silica gels 389

Polyglycidyl methacrylate chains 710

Polyglycidylmethacrylate grafted-tentacle gel matrix 715

Polyhydroxymethacrylate 106

Polymethacrylate 706

Polymyxin B sulfate 561

Polynucleotide phosphorylase 432

Polyols 38

Polyoma virus medium size tumor antigen 537

Polyoxyethylene alkyl ether 530

Polypeptide 141

Polysaccharides 620

PolySep 106

Polystyrene 673

Polystyrol 706

Polyvinyl pyrrolidone 623

Porath 310

Porcine growth hormone 185

Pore size

buffers and additives 860

cell affinity chromatography 669, 680

chromatofocusing 398

chromatography on porous matrices 787–788, 791

coupling reactions 709

covalent chromatography 644

displacement chromatography 518

dye-ligand affinity chromatography 417

hydroxyapatite chromatography 280, 285

immobilized metal ion affinity chromatography 313–316

integral membrane proteins 546

ion exchange chromatography 8, 27, 77

liquid-liquid partition chromatography 472–473, 486

non-porous media 786

recombinant proteins 577

reversed-phase chromatography 156, 158, 160, 168, 172, 175, 177–179, 184–186, 203–204, 206, 210–212, 214–216, 218, 220, 222

scale-up of downstream processing 775, 781

size exclusion chromatography 94–95, 112–113, 117, 128, 143

Pore size distribution 143, 791

POROS 50, 775, 787, 791, 807

POROS gels 787, 789–809

POROS HS 820

POROS ion exchangers 55

POROS R2 819

POROS S 820

Porosity 6, 16, 24, 26, 32, 79, 92, 128, 176, 313, 646, 714, 735, 749, 755, 758

Porous glass 113

Porous matrices

see Contents 785–786

advantages and disadvantages 788

antibody purification and screening 823

application examples 813–823

back pressure 788, 798, 806, 811

binding capacity 788

column dimensions 808

column packing 800

equipment 789

flow rates 788–790

gels on porus matrices 791, 798

general utility 788

optimization 810

particle sizes 809

pressure 799

sample preparation 800

scaling up and down 808

storage 812

troubleshooting 824

work stations 810

Positive selection 678–679, 694

Post-run analysis 866

Post-translational modifications 559

Potassium chloride 405

Pre-bleeding 323, 338, 353

Pre-charging 323

Precipitation effect 850

Pre-column 154

Pre-packed column 280

Preparative columns 290

Preparative-sized column 463

Preparing a buffer 846

PreScission protease 563

Presence 854

Preservatives 59

Pressure 799

Pressure drop 687, 692

Pressure variations 94

Price(s)

see Cost(s)

Primary amines 321, 851

Principles 706

Probond 566, 592

Processing capacity 685, 690–691

Procion Brown MX-5BR 428

Procion Green H-4G 428, 436

Procion Navy H-ER 436

Procion Red 486

Procion Red H-E3B 428

Procion Red MX-2B 436

Procion Yellow MX-8G 436

Procion Yellow MX-GR 436

Procion^® dyes 428

Procollagenase 435

Proinsulin 308, 358

Prokaryotes 767

Promoter 588

Promoter operator 591–593

Properdin 734

PROSEP-Heparin 744

Prostaglandin H₂ synthase 535

ProSys 777

Protease inhibitors 41, 291, 306, 337, 623, 750, 827, 841, 854, 856

Proteasome 558

Protecting 841

Protein A 365, 563, 672–673, 678, 706, 718, 772, 798

Protein A affinity resins 772

Protein A-Sepharose 633

Protein binding 806

Protein binding capacity 709

Protein bioactivity 826

Protein degradation 404

Protein design 138

Protein determination 247, 262

Protein folding 826

Protein G 365, 718, 798

Protein KW 101

Protein PAK 60 100

Protein splicing 560

Protein structure 826

Protein synthesis factors 733

Protein test mixture 285

Protein tyrosine phosphatase 536

Protein-DNA interactions 587

Proteoglycans 354, 629, 742

Proteoglycan core proteins 622

Proteoliposomes 140

Proteolytic degradation 582–583

Proteolytic enzymes 131

PS I- and II-monomers 297

PS I-trimer 297

Pseudo-bioaffinity ligands 707

Pseudomonas aeruginosa 264

Pulsation 94

Pulse damper 129

Pulse detergent gradient 453, 455

Pulse gradient 454

Purification by HIC 264

Purification tag 590

Purity 846

Pyrogen testing 693

Pyroglutamyl aminopeptidase 653–654

Pyrrolidonecarboxylyl peptidase 653

Rat lens 535

Rat liver microsomes 41

Reaction center from Rhodopseudomonas spheroides 535

Reaction center Photosystem I and II 535

Reaction intermediates 816

Reactive dyes 426–427

Reciprocating pistons 94

Recombinant

chromatofocusing 407–410

chromatography on porous matrices 813, 815–816

chromatography using Strep-tag affinity peptides 826–827, 830–833

DNA–binding proteins 588, 597

hydrophobic interaction chromatography 235, 238

immobilization of nucleotides 719, 726

immobilized metal ion affinity chromatography 306–308, 324, 335, 337, 357, 360, 363–364, 368–369, 377

miscellaneous biospecific affinity ligands 739, 747, 754, 755, 759

recombinant proteins 557–586

reversed-phase chromatography 208–209, 215–217, 228–229

scale-up of downstream processing 766, 773

size exclusion chromatography 137–138, 150

Recombinant peptides and proteins 215

Recombinant protein production 826

Recombinant proteins

see Contents 557

application examples 569–579

contaminating proteins co-eluting with the fusion protein 582

disulfide bonds 571

expression systems 559

failure to bind to affinity resins 583

formation of inclusion bodies 579

fusion partners 559–560, 562–563, 568

fusion proteins 306, 308

gel matrices 564, 566–567

his-tagged proteins 564

on-column cleavage 576

sample preparation 561

troubleshooting 579

usefulness of HIC 237

Recombinant Tick anticoagulant peptide 813

Recovery 275

Red blood cells 463

Redox titrations 140

Reductase 754

Reductive amination 626–627

Refraction detector 513

Regeneration protocol 345

Regeneration

see Gel(s), regeneration

Regulatory support file 706

Remazol^® dyes 428

Renaturation 560

Renex 287

Replenine 371

Repressor gene 591–593

Repressor purification 589

Reproducibility 454, 457

Reservoir 11

Resin(s)

see Gel(s)

Resolution capability 409

Resolution 80, 275, 404, 788, 808

Respiratory syncytial virus 537

Restriction endonuclease 308, 360

Retinol-binding protein 308

Reverse transcriptase 306

Reversed IMAC 372

Reversed-phase 517

Reversed-phase chromatography

see Contents 147–148

alkyl chain length 177–178

angiotensin II 193

application examples 195–223

carbohydrate microheterogeneities 217

column conditioning 187

column lifetime 191

column-ovens 155

columns 157

desalting 217

desalting in reversed-phase 217

detectors for reversed-phase 154–155

endcapping 174

flow rates 183

glycoproteins 220

hydrophobicity 149

large proteins 218

MALDI-MS 155

membrane proteins 222

optimization 188

particle size 178

pore size 178

preparation of eluents 185–187

principle 149–153

recombinant peptides and proteins 215, 217

removal of detergents 217, 223

reversed-phase column cleaning 190

seoaration of prtoeins 450

silicas 174

stationary phase 156–180

trace enrichment 217

troubleshooting 194, 196

Reversed-phase HPLC 545

Rhinovirus receptor 537

Rhodopsin (large hydrophobic peptides) 538

Ribonuclease A 252, 285–286

Ribonuclease 306, 401

Ribosomal proteins 202

Ribosyl moiety 720

Ricinus communis 620

Ricinus communis agglutinin 629, 633–634

Rigid hydrophilic polymer 109

RNA polymerase 579, 588, 608, 612–614, 739, 741

RNA polymerase holoenzyme 608

RNaseA 826

Rosetting 669, 679, 695

RPC

see Reversed-phase chromatography

Saccharomyces cerevisiae 305, 739

Salts 291

Salt gradients 454

Salting out 114

Salting-in effect 850

Salting-out effect 850

Sample application 411

Sample loading limits 188

Sample loop 13, 94, 282

Sample preparation, for

chromatofocusing 398

chromatography on porous matrices 800

covalent chromatography 737, 739, 741–742, 750, 755, 759

dye-ligand affinity chromatography 437

hydroxyapatite chromatography 275, 280

immobilized artificial membrane chromatography 451

immobilized metal ion affinity chomatogr. 319, 325–326, 330, 332, 336

ion exchange chromatography 41

liquid-liquid partition chromatography 479

recombinant proteins 561, 641, 646, 652–653

reversed-phase chromatography 148, 155, 187

size exclusion chromatography 116

Sample rotator 621

Sample viscosity 741

Sandwich formation 634

Saralasin 193

Sawtooth gradient 403, 811

Scale-up of downstream processing

see Contents 765

buffers for large scale application 774

column bed volumes 780

column dimensions 780

for large-scale applications 772, 774–775

microfiltration 767

price(s) 774

scale-up 765, 776

selection of chromatographical methods 771

Scale-up of downstream processing 765

Scaling down 57, 342, 486–487, 690, 766, 808

Scaling up 54, 56–57, 124, 254, 310, 342, 364, 406, 486–487, 652, 690, 694, 765–767, 769–770, 773–777, 779, 781–782, 788, 808–809, 813, 820, 822, 831, 833, 872, 875

see also Large scale

Scanning densitometry 459

Scouting run function 872

Screening of dye-ligands 442–443

SDS 127, 131, 288, 852–853

SDS-PAGE 452, 842

SEC

see Size exclusion chromatography

Secondary amine 708, 851

Selectins 622, 845

Selectivity 511

Self-aggregation 852

Self pack device 801

Self Pack 803

Self-packing

see Column(s), packing of

Semliki Forest virus 533

Sendai virus 531, 535–538

Separation criteria 386

Separation of monomers and dimers 491

Sephacryl S 103, 200 656

Sephacryl 104, 775

Sephadex G-25, 200, 648, 673

Sephadex 620, 775

Sepharose 313, 626, 775

Sepharose 4B 673

Sepharose 6B 673

Sepharose 6MB 672–673

Sepharose CL-2B 623

Sepharose Fast Flow 313

Sepharose High Performance 313

Sepharose HP 775

Sequencing 842

Serine proteases 855

Serine-dependent proteases 354

Serum albumin 491

Serum proteins 306, 733

Serum 364, 490

Sex hormone-binding globulin 490

Shape 120

SHBG 489

Shear rate 686

Sialic acid 631–632

Sialyltransferase 726

Silane 859

Silanol groups 114

Silica subsurface amines 461

Silica 117, 174, 314, 517

Silica-based gels 313

Site shadowing 685

Size exclusion 93, 112, 849

Size exclusion chromatography

see Contents 89–90

application examples 134–143

buffer exchange 95

calibration 94

choice of buffer 114

column and gel maintenance 129–133

column equilibration 121

delipidation of membrane proteins 140–141

desalting 90

detergent 94, 120, 123, 136

detergent exchange 140

elution modes in size exclusion 125, 127

equipment for size exclusion 93

fast desalting 105

gels for gel filtration, gel permeation and size exclusion 91, 112–113, 787

LALLS photometer 137

molecular weight determination 134–137

optimization 122–125

packing the column 118

sample loading, washing, elution 122

sample preparation 116–117

storage of size exclusion column 133

system set-up 116

troubleshooting 134–135

UV detector 93, 137

void volume 91

Size exclusion HLPC 537

Sjögren's syndrome 497

Slurry 11

Sluyterman 387

Small zone 138

SMART system 117, 325

sn-1.2-diacylglycerol kinase 535

Snake PLA₂ 457–458

SNAP 33, 56, 74, 80

Socket valves 94

Sodium acetate 852

Sodium ascorbate 409, 857

Sodium azide 133, 292, 693, 861

Sodium chloride 405, 851

Sodium cholate 65, 288, 530

Sodium deoxycholate 530

Sodium dodecylsulfate 321, 529–530

Sodium perchlorate 183

Sodium taurodeoxycholate 530

Soft release avidin 591

Software 11, 15, 451, 681, 740, 810

see also Computer chromatography software 863–876

Software packages 868

Software updates 875

Software validation protocol 875

Solenoid valves 15

Solid phase syntheses 726

Solubility 386

Solubilization 529

Solubilization buffer 465

Solubilizing 464, 841

Soluble amphiphiles 852

Solute 5

Sonication 831

SOTA phase 112

Source 30, 775

SOURCE 787, 791, 807

Soybean lectin 684

Spacer arm 688

Spacer displacers 515

Spacer molecules 417

Spacer technologies 709

Spacer 336, 707, 723

Specific resolution 112

Specific site 606

Spectral properties 852

Speed-vac 217

S-peptide 562

Sperm–egg recognition 632

Splenocytes 691

S-protein 826

S-protein agarose 566

Stacking effects 428

S-tag 826

Standard run 800

Standards for HPSEC 119

Stationary phase(s)

see also Gel(s)

chromatofocusing 389–390, 392, 402, 406–407

covalent chromatography 640

displacement chromatography 506, 508–511, 513, 515–517

hydrophobic interaction chromatography 257

immobilized artificial membrane chromatography 450, 454

immobilized metal ion affinity chromatography 304

ion exchange chromatography 5–11, 13, 15–17, 22–29, 31–33, 38–41, 43, 45, 47, 52, 54–60, 62, 64–67, 69, 71, 74–80

liquid–liquid partition chromatography 471, 475, 478, 481, 484, 488–489, 502

reversed-phase chromatography 149–154, 156, 175–177, 188, 199, 202

size exclusion chromatography 91, 95, 114, 128

Steel 790

Stem cells 676, 678

Stem cell isolation 695

Step gradient detergent elution 453

Step gradient 288, 809

Steric accessibility 252

Steric anion exchange chromatography 273

Sterile environment 692

Steroid receptors 733

Stokes radius 118, 141

Storage 812

Strep-peptide 562, 565

StrepTactin 830

Strep-tag 830, 833

Strep-tag II 830

Strep-tag affinity peptides

see also Contents 825

membrane protein complexes 835

metalloproteins 827

practical performance 830–833

scale-up 833

StrepTactin affinity columns 831

troubleshooting 834–835

Streptavidin 562–563, 565–566, 568, 591, 595, 600, 827, 829–830, 833–834

Streptavidin tag 591

Streptavidin-agarose 566

Streptococcus suis 658

Stromelysin (metalloendopeptidase) 434

Strong ion exchanger 80

Submandibular and parotid gland (rat) 536

Submaxillary mucin 631–632

Substance P neuropeptide receptor 537

Substance P receptor 543

Substrate composition 841

Subtilisin 309

Sucrose density gradient 297

Sucrose synthase 309

Sucrose 115, 859

Sugar phosphate transporter (E. coli) 537

Sugars 720

Sugar-specific antibodies 620

Sulfations 627

Sulfhydryl blocking agents 654

Sulfhydryl reagents 291

Sulfhydryl reagents 40

Sulfobetain 530

Sulfobetaines 529

Sulfone bridges 707

Sulfone-activated sepharose 626

Sulphonated polystyrenes 118

Sulphur 237

Sumner 620

Supercomplexes 816

Superdex 200 478, 483

Superdex Peptide 101

Superdex 103, 117, 139, 775

Superflow 316

Superformance 779, 802

Superloop 13, 282, 791

Superose 102, 127, 313

Superoxide dismutase 306, 734

Superparamagnetic beads 373

Suppliers 887–894

Support 473, 875

Surface active agents 852

Surface chemistry 807

Surface hydrophobicity 237

Surface properties 498

Surface topography 309

Surfactants 852

Swelling behavior 23

SynChropak GPC 107

Synechocystis 814, 295

Syntheses

chromatography on porous matrices 791, 818, 819

coupling reactions 706, 708, 710–715

covalent chromatography 643, 650, 659

DNA-binding proteins 588, 595, 602

hydrophobic interaction chromatography 239, 241, 252

immobilization of nucleotides 723–724, 726–727

immobilized metal ion affinity chromatography 313, 336, 348, 363, 373

miscellaneous biospecific affinity ligands 733, 739, 742, 745–747, 752

reversed-phase chromatography 156–174

Syringe 282

Syringe operation 330

T helper 694

T7 peptide 563

T8 antigen 533

Tailing 507

Talon 566

Tandem column 334

Tandem 323

Tannins 623

Taurine 280, 291, 395

Taurodeoxycholate 529

TCA precipitate 453

TCA precipitation 453

TEA 217

TED 304, 306, 309, 311–312, 316–317, 334–335, 350–352, 372, 374, 622, 632, 690

TED-Agarose 316

Temperature effects 853

Temperature

buffers and additives 845, 847, 850, 852, 853

cell affinity chromatography 670, 674, 682, 684, 693, 697

chromatofocusing 389, 395–396

chromatography on porous matrices 806, 811–812, 814

chromatography software 872

chromatography using Strep-tag affinity peptides 827

coupling reactions 714–715

covalent chromatography 643, 646, 658, 663

displacement chromatography 516–517

DNA-binding proteins 595, 597, 602, 608

dye-ligand affinity chromatography 428–429, 436, 438–439

hydrophobic interaction chromatography 239, 245–246, 248, 256, 267

hydroxyapatite chromatography 275, 279, 284, 289–290, 292

immobilized artificial membrane chromatography 451–452

immobilized metal ion affinity chromatography 321, 325, 334, 336, 339, 353

integral membrane proteins 532

ion exchange chromatography 11, 33, 44, 46, 49, 51, 66

lectins and glycoconjugates 621

liquid-liquid partition chromatography 472, 474–475, 477, 480–483, 488

miscellaneous biospecific affinity ligands 734, 743, 748, 750, 758

recombinant proteins 560, 576, 582

reversed-phase chromatography 155, 178, 183–185, 188–190

scale-up of downstream processing 769

size exclusion chromatography 93, 118, 122, 125, 130, 132–134, 137, 142

Template chromatography 742

Tensides 852

Tentacle ion exchangers 7, 55

Tentacles 113, 313

TEPA 344

Test mixture 284

Test separation 400

Tetanus toxin 309

Tetrabutylammonium salts 182

Tetracyclines 307, 372

Tetraethylenepentamine 344, 364

Tetrahydrofolate reductase 433

Tetrahydrofuran 629

TFA 153, 181–182, 217

T-helper cells 678–679, 688

Theoretical plates 122, 128, 808

Theoretical titration curve 843

Thesit 530

THF 304, 306, 309, 311–312, 316–317, 334–335, 350–352, 372, 374, 622, 632, 690

Thimerosal 652, 751, 861

Thiobond 567

Thiobond Resin 593

Thioether bridges 707

Thiol (SH-) reagents 854

Thiol covalent chromatography 640–641

Thiol functional group 641

Thiol oxidation 651

Thiol proteases 855

Thiophilic adsorption 707

Thiophilic adsorption chromatography 237

Thiopropyl Sepharose 648, 659

Thiopropyl Sepharose 6B 645

2-Thiopyridone 242, 648

Thioredoxin 309, 563, 593

Thrombin 590

Thrombin^b 563

Through-flow pores 787

Thylakoid membrane 294, 814

Thymidine kinase 308

Thymocytes 677

Thyroglobulin 487

Tick-borne encephalitis virus 535, 538

Tissue lectin 630

Titration 396

Titration curve 842–843

TLCK 856

TLPK 856

T-lymphocyte 694

TNB-thiol gel 645

Topoisomerase II 739

Toxic 39, 41, 317, 475, 589, 592, 594, 625, 714, 720, 834, 854–855, 859

Toyopearl 108

Toyopearl HW 113

Trace enrichment 217, 339, 346

Training courses 875

Transferases 752

Transferrin 363, 366, 487, 489–490, 499

Transmembrane pressure 768

Treatment of glass surfaces with silane 859

TREN-Novarose gel 365

Trichlorbutanol 861

Trichloroacetic 452

Triethylamine 621

Triethylammonium acetate 183, 186

Trifluoracetic acid 132, 153

Triglyceride lipase 734

Trimethylamine 852

Trimethylchlorosilane 174

Tri-n-butyl phosphate 370

Tris 392

Tris (hydroxymethyl) amine 672

Tris(2-aminoethyl)amine 335, 364

Tris(carboxymethyl)ethylenediamine

see TED

Trisacryl 673

Triton 273, 287, 853

Triton X-100^® 116, 140, 529–531, 652, 751

Triton X-114 116, 530

TRIZMA 45

tRNA methyltransferase 739

Tropomyosin 52

Troubleshooting, for

cell affinity chromatography 696–677

chromatofocusing 413

chromatography on porous matrices 824

chromatography using Strep-tag affinity peptides 834–835

covalent chromatography 662–663

displacement chromatography 522

dye-ligand affinity chromatography 444–445

hydrophobic interaction chromatography 257–258, 266–267

hydroxyapatite chromatography 296–297

immobilized metal ion affinity chromatography 374–377

integral membrane proteins 548

ion exchange chromatography 77–78

lectins and glycoconjugates 634–635

liquid–liquid partition chromatography 501–502

miscellaneous biospecific affinity ligands 759–761

recombinant proteins 579

reversed-phase chromatography 194, 196

size exclusion chromatography 134–136

Tryglycylglycine 688

Trypan blue 671

Trypsin 135

Trypsin inhibitors 306

Trypsinogen 141

Tryptophan 154

TSK 4000SW 124

TSK Toyopearl 775

TSK-gel G3000SW 113

TSKgel PW series 112

TSKgel SW type series 112

TSK-PW-type 127

TSK-SW-type 127

T-suppressor cells 678–679, 688

Tuna 304

Tween 852–853

Tween 20 410

Tween 80 530

Two-hybrid system 588

Two-phase system 474, 532

Two-step detergent gradients 459

Two-step linear gradient 462

Ubiquitin 308–309

UDP 718

UDP-glucose dehydrogenase 430

UDP-glucoseceramide glycosyltransferase 435

UDP-glucuronosyltransferase 535

Ultrafiltration 217, 623, 646, 767, 860

Ultrasonic disintegration 561

Ultrogel 626

Ultrogel AcA 44 631–632

UMP 718

Unbound proteins 282

Unfolding 141

Unfolding of protein 238

Unspecific interactions 23

Unwanted ionic interactions 849

Uracil 193

Urea 38, 127, 141, 292, 314, 405, 743, 858–859

Urtica dioica lectin 620

UTP 718

UV detector 93

UV spectrometer 137

UV/VIS-monitors 13

UV-detection 154

UV-monitor 12

UV-transparency 321, 533

van Deemter 30, 80

van Deemter equation 80

van Deemter plot 30

Vantage A, S 779

Variable wavelength 13

Varicella zoster virus 537

V _e 91

V_H domains 367, 828

V_L domains 367

V _o 91

Virgil 72, 73

Viscometers 143

Viscosity 51, 116, 849

Viscous solution 282

Viscum album 624–625

Viscum album agglutinin 624

Vitamin B₁₂ 193

Void volume 91, 282, 680–681

Volatile buffers 280, 846

Volatile ion-pair reagents 217

Volatile mobile phases 150

Washing 454

Water for injection 692

Weak anion exchanger 387

Weak ion exchanger 81

Western blotting 842

Wheat germ agglutinin 628, 673

Wide bore tubing 118

Work stations 810

X-ray 814

Xylose isomerase 308

Yeast cell extracts 259

YMC–Pack Diol 101

Z domain 563

Zirconia 78

Zirconium dioxide 78

Zone widening 409

Zorbax Bio–Series 97

Zorbax GF-250 95

Zorbax PSM 60 98

Zwitter ionic detergents 405

Zwittergent 530

Zwitterionic 845

Zwitterions 40