Index

Note: Figures and Tables are indicated by italic page numbers, Boxes by emboldened numbers absorption/distribution/metabolism/excretion AIDS (ADME) studies 74–5 deaths due to 408 Actimmune 224, 232 gene therapy 423, 444–5 Activase 346 AIDS vaccine activin(s) 311, 315, 317–19 clinical trials 409–10 adeno-associated virus, as gene vector 429 development of 396, 407–8 adenosine deaminase (ADA) defi ciency diseases, diffi culties associated 409 treatment of 440 alanine 15, 16, 165 adenoviruses albumin 5, 354–5 as gene vectors 428–9 as excipient 164, 350 manufacture of 431 Aldurazyme 362 as vaccine vectors 406 Alfatronol 224 adjuvants 412–16 allantoin 361, 362 bacteria/bacterial products as 412, 414–15 Alteplase 348 mineral-based 412, 413–14 aluminium-absorbed vaccines 413 mode of action 413 aluminium-based adjuvants 412, 413–14 oil-based emulsions as 412, 414 Amgen (biopharmaceutical company) 7 adult stem cells 459–60 amino acid analysis, characterization advantages compared with embryonic stem cells 460 by 185–6 see also haematopoietic stem cells amino acids 15, 15, 17 Advexin 442 bonding between 16, 18–19 affi nity chromatography 140, 141, 148–50 charged (at pH 7) 15, 17, 142, applications 133, 140, 150,COPYRIGHTED 158, 337, 351 144,MATERIAL 145 contamination of product by 199 deamidation of 17, 161, 162 affi nity-based purifi cation of mRNA 44–5 hydrophobic 17, 146–7, 147 agarose gels, in hydrophobic interaction modifi ed 17 chromatography 147–8 as stabilizing agents 164–5 Agrobacterium-based vector-mediated gene structure(s) 16, 145 transfer 116 α-amylase 365

Pharmaceutical biotechnology: concepts and applications Gary Walsh © 2007 John Wiley & Sons, Ltd ISBN 978 0 470 01244 4 (HB) 978 0 470 01245 1 (PB) 466 INDEX anaemia aptamer(s) 8, 10, 453 causes 275, 278 Aranesp 7, 8, 266, 276 treatment of 266, 274–7 arcitumomab 380, 384 anaphylactic shock 350 arginine 15, 16, 17, 144, 145 ancrod 5, 341 armyworm (Spodoptera frugiperda), vaccines androgens 315, 316 produced in 118, 119 biological actions 315, 316–17 ‘artifi cial ’ (closed-loop infusion pump animal cell culture systems 127–9 system) 305 bioreactor design 126, 128 aseptic fi lling 166 as source of biopharmaceuticals 109–10 validation of 201 anion exchange 144, 145–6, 146 asparaginase 355–6, 357 anion-exchange chromatography 145–6, 146, 158 asparagine 15, 16, 17 anthrax vaccines 397, 400 deamidation of 17, 161, 162 antibiotics, assay method for 176 synthesis of 356, 357 antibodies aspartic acid/aspartate 15, 16, 17, 142, 145, 161, 162, as diagnostic reagents 177–9 357 virus-specifi c 198 Atryn 115, 345 antibody–antigen binding 378, 379 autoclaves, validation of 201 antibody architecture. 382–3 autoimmune conditions 395 antibody fragments 394 Avastin 381, 394 antibody libraries, production and screening Avonex 7, 8, 224, 230 of 377–8 antibody-directed enzyme prodrug therapy B-lymphocytes 207 (ADEPT) 386–7, 388 activation by 252 antibody-mediated approach to tumour detection/ baby hamster (BHK) cells 8, 109 destruction 383–6 Bacillus subtilis, proteolytic enzymes 364 limitations 388–9, 394 bacterial artifi cial (BACs) 48 reasons for clinical disappointment 388–9 bacterial products, as adjuvants 412, 414–15 antibody-mediated cell targeting 378, 379 bacteriophage λ 48 anticoagulants 340–5 basophils 207 antihaemophilic factor 330 Bayovac CSF E2 9, 411 α 2-antiplasmin 352 BCG vaccine 397, 398 antisense oligonucleotides 445–6 BCL-2 oncogene products 448 advantages 448 bead milling, cell disruption by 135–6 delivery systems 450–1 266 disadvantages 448 BeneFix 335, 339 manufacture of 451 Beromun 260, 261 mode of action 446–7 Betaferon 224, 230, 231 modifi cations 449 Betaseron 224, 230 pharmacokinetics 450 381, 394 phosphorothioate-based 449 bicinchonic acid-based protein assay method 179, uses 448 180 antisense technology 8, 10, 445–50 bioassays overview 446 drawbacks 176–7 antisera 372, 374 product potency determined by 176–7 production of 372, 373 viral assays 198 antithrombin 341, 344–5 bioavailability studies 75 recombinant 115, 345 bioequivalence studies 75 α 1-antitrypsin 353–4 Biogen (biopharmaceutical company) 7 Apidra ( glulisine) 298 biologically sourced products 1, 2 Apligraf 457 biologic(s) 1, 371 INDEX 467 biologics licence application (BLA) 92 bone remodelling, role of IGFs 281 biopanning 376 bovine milk 114 biopharmaceutical companies 6–7 ‘breathing’, in protein structures 28 biopharmaceutical(s) brushite 154 current status 8, 9 buffer 131 future prospects 9–11 growth of industry 3–7 CA125 390 meaning of term 2 cachectin see tumour necrosis factor α sources 105–20 324–5 biospecifi c affi nity chromatography 149–50 calicheamicin (cytotoxic antibiotic) 385, 386 biotechnology cancer meaning of term 1–2 chemotherapy 278 patenting in 68–70 gene therapy 441–4 Biuret protein assay method 179 treatment with interleukins 246–8 blastocysts, recovery of inner cell mass 457–8 cancer vaccines 410–11 blood, transgenic animals, as source of therapeutic capillary electrophoresis 182 proteins 115 carbohydrates, as stabilizing agents 165 blood cells 267 carcinoembryonic antigen (CEA) 390 see also leukocytes see also CEA-SCAN blood coagulation pathway 330–2 carcinogenicity tests 83, 84, 85 terminal steps 332–4 CaroRX 118 blood factor I 330, 332, 333, 334 Carticel 456 blood factor II 330, 332, 333 cartilage 456 blood factor III 330–1, 330 types 456 blood factor IV 330 cation exchange 144, 145, 146 blood factor V 330 cation-exchange chromatography 144, 158 blood factor VII 330, 330, 331 cationic lipids 433 blood factor VIIa, recombinant 340 interaction with DNA 434 blood factor VIII 330, 332, 335 CD4 antigen 407, 408, 444 defi ciency 334, 335–6 CEA-SCAN 380, 384 production of 336–9 production of 385 purifi cation of 140, 150, 336–7 cell banking systems 121–3 recombinant 150, 335, 336, 337–8, 338–9 cell culture systems 127–9 possible contaminants 338 cell disruption procedures 134–6 blood factor IX 115, 330, 332 cell-based therapies 453–60 defi ciency 339 cellulase 365 production of 339–40 Center for Biologics Evaluation and Research recombinant 339 (CBER) 90, 91–2 blood factor X 330, 331, 332 product types covered by 92 blood factor XI 330, 332 Center for Devices and Radiological Health blood factor XII 330, 331, 332 (CDRN) 95 blood factor XIII 330 Center for Drug Evaluation and Research defi ciency 340 (CDER) 90, 91–2 blood factors product types covered by 92 listed 330 centrifugation 134, 136 notation 330 Ceredase 360 blood products 5, 6, 9, 335–40 Cerezyme 360 production of 336–7, 456 CGD, treatment of 232 blood transfusion 455–6 chaperones, inclusion body formation and 10 blue dextran 152 chemical-based drugs 1, 2, 3 bond energies 27 chickenpox vaccines 398, 399 468 INDEX chimaeric antibodies 392–3 pyrogenic 189–91 examples (listed) 380, 381 detection of 191–5 Chinese hamster (CHO) cells 8, 109 viral 197 interferons produced in 230 Coomassie Blue dye, protein assays using 179, 180 chondrocytes 456 coronary thrombosis 340 choriogonadotrophin alfa 322 covalent bonding, in proteins 28 Christmas disease 339 Creutzfeldt–Jakob disease (CJD) 309–10 Christmas factor 330 cryoprotectants 168, 438 chromatin 41 cyclosporin A 249 chromatofocusing 141, 155 cysteine 15, 16, 17 chromatographic purifi cation 140–55 oxidation of 17, 162, 163 columns 140–1, 143 cystic fi brosis 357–8, 441 chromosomes 41 clinical symptoms 357–8 number in various species 42 genetic basis 358 chymotrypsin 14, 364 management of symptoms 358 clean-room conditions 131, 133 treatment by DNase 358–9 validation of 201 treatment by gene therapy 358, 423, 440–1 cleaning, decontamination and sanitation (CDS) cystic fi brosis transmembrane conductance regulator procedures, validation of 202 (CFTR) gene 358, 423, 440 clinical trials 57, 84, 86–9 receptors 210–11 design 87, 88 59, 205–63 phases 86–7 as adjuvants 415 clotting disorders 334–5 as biopharmaceuticals 211–12 clotting factors classifi cation 205, 208 listed 330 developments in understanding of biology 208 notation 330 generalized characteristics 208–9 see also blood factor listed 206 clusterin gene 448 cytotoxic antibiotic(s) 385, 386 coagulation pathway 330–2 cytotoxic T cells 247, 248 terminal steps 332–4 collagenase 356, 364 darbepoetin alfa 266, 276 colony-stimulating factors (CSFs) 5, 269–72 debriding agents 364 cells targeted by 266, 267 delivery systems 70–4 clinical applications 270–2 protein immunogenicity caused by 78 clinically approved products 266 2Ј-deoxyribose 38, 39 molecular mass 195, 269, 270 see also DNA column chromatography, meaning of term 140 mellitus 292 complementarity-determining regions diafi ltration 139–40 (CDRs) 382 diagnostic imaging, monoclonal antibodies complementary DNA (cDNA) 37 used 378 cloning 51 dicoumarol 341 concentrating crude protein product 137–40 digestive aids, enzymes as 364–6 conjugated proteins 13–14 diphtheria vaccines 397, 399–400 contaminant-clearance procedures 166 disruption of cells and tissues 134–6 validation of 201 chemical methods 134, 135 contaminants mechanical methods 134–6 detection of, immunological approaches 185–9 disulfi de bonds DNA-based 195–6 albumin 354, 355 microbial 196–7 HIV 407 protein-based 173–4 disulfi de exchange (in polypeptides) 162, 163 detection of 180–5 disulfi de interchain links (insulin) 293, 297 INDEX 469 disulfi de intrachain links embryonic stem cells 457–8 blood-clotting factors 334 differentiation of 459 growth 307 Enbrel 7, 260 immunoglobulins 382 endocytosis, receptor-mediated 76, 77, 435, 450 DNA 38 endotoxin 189, 191 complementarity in 40–1 medical reactions 190, 191 structure 38, 40–1, 41 molecular structure 191, 192 DNA arrays 61–2 removal of 195 DNA-based contaminants 195–6 Engerix-B 401, 402, 405 DNA cloning 47–54 engineered biopharmaceuticals expression vectors 8, 47, 53 298, 301–4 polymerase chain reaction used 51–3 tissue plasminogen activators 6, 346, 348–9 DNA hybridization studies 196 enteric-coated products 71 DNA removal (from product) 196 enzyme immunoassay (EIA) systems 177–8 validation of 201–2 enzyme-linked immunoassay (ELISA) system 178 DNase 70, 356, 357–9 enzymes 355–66 dopaminergic , differentiation from as digestive aids 364–6 embryonic stem cells 458, 459 eosinophils 207 ‘dot blot’ assays 196, 198 epidermal (EGF) 282–3 downstream processing 120–1, 131–72 biological activities 282 contaminant-clearance procedures 166 cells targeted by 266, 282 validation of 201 medical applications 267 overview 132 molecular mass 195 drotrecogin alfa 345 epoetin alfa 266 drug approval systems epoetin beta 266, 276 European Union 96–100 Epogen 7, 8, 266 Japan 100 Eprex 78, 164 USA 92–5 Epstein–Barr virus 391–2 world harmonization of 101 eptacog alfa-activated 340 drug development 57–104 (EPO) 3, 272–8 costs 7 assays for 176 timescale(s) 7, 86 biological activities 209, 211, 274 drug discovery 57, 58–9 cells targeted by 266, 267 genomics approach 59–61 dosage/activity 276 drug-based tumour immunology 386–7 molecular mass 195, 273 Dukoral 401, 402 non-renal applications 277, 278 Dulbecco’s modifi ed Eagle’s medium 127 sales data 8, 275 dye affi nity chromatography 152 structure 14 applications 120 therapeutic applications 266, 274–7 disadvantages 152 three-dimensional structure 273 dyes as ligands 152, 153 Eschericheria coli (E. coli) interferons produced in 226, 227, 228, 230, 231 Ecokinase 346, 348 lipopolysaccharide on outer membrane 107, 109, 191 production of 348–9, 349 recombinant proteins produced in 8, 105–9 Edman degradation method 19, 188, 189 advantages 106 EGF receptor 282 drawbacks 106–7 see also use in DNA cloning 47, 48, 50 eIF-2α protein kinase system 221–4 etoposide 387, 388 electrostatic interactions, in proteins 27 eukaryotes embolism 340 chromosomes 41 Embrel 8 genes 43, 44 470 INDEX

Europe, drug registration in 95–100 Food and Drug Administration (FDA) 90–5 European Commission 96 organizational structure 91 European Medicines Agency (EMEA) 96–100 responsibilities 91 Committees 97–8 substances regulated by 90 marketing authorization on toxicology tests 82 centralized procedure 98–9 Food, Drug and Cosmetic (FD&C) Act 90 mutual recognition procedure 100 Forsteo/Forteo 324 objectives 98 freeze drying 168–9, 170–1, 438 structure and responsibilities 97 Freund’s complete/incomplete adjuvants European Patent Directive 69–70 (FCA/FIA) 412, 414 European regulations 95–100 functional genomics, approaches 60–5 excipients 164–6 fungi, as source of biopharmaceuticals 111 expression vectors (gene cloning) 8, 47, 53 fused proteins 6 Exubera (inhalable insulin) 11, 71, 298, 304 inclusion body formation and 108 -2 in 250–1 Fabrazyme 360 Fabry disease, treatment of 360 α-galactosidase 356, 360–1, 365 factor see blood factor β-galactosidase 366 familial hypercholesterolaemia, treatment by gene Gaucher’s disease 359 therapy 440 treatment of 359, 360 fast protein liquid chromatography gel-fi ltration chromatography (FPLC) 157 downstream processing by 140, 141, 142 Fasturtec 363 endotoxin removal by 195 female follicular structure 313, 314 GEM 21S 266 female reproductive cycle 312 Gendicine 10, 419, 442 in 312, 318 gene chip arrays 61–2 see also follicle-stimulating hormone; luteinizing gene cloning 47–54 hormone expression vectors 8, 47, 53 fermentation process polymerase chain reaction used 51–3 industrial-scale 123 gene neighbourhood method 61 microbial cell fermentation 124–7 gene sequence, amino acid sequence determined α-fetoprotein (AFP) 390 from 21 fi brin 330, 332, 333, 347 gene therapy 10, 419–24 fi brin-stabilizing factor 330 and AIDS 423, 444–5 fi brinogen 330, 332, 333, 334 basic approach 420–3 fi brinokinase see tissue plasminogen activator and cancer 441–4 fi brinolysis 345 and cystic fi brosis 358, 423, 440–1 fi broblast growth factors (FGFs) 266, 284 ethical considerations 445 medical applications 7, 106, 266, 285 factors affecting choice of protocol 423–4 see also keratinocyte growth factor fi rst trial 420 fi lgrastim 266, 271, 272 and genetic disease 438–41 fi lling process 166–7 in situ approach 422, 423 fi nal product fi ll 166–7 in vitro approach 422, 423 fi nal product formulations 159–71 in vivo approach 422, 423 stabilizing excipients in 164–6 plasmid DNA as vector 432–6 fl ash freezing, plasmid DNA products 438 principles 420 ‘fl atulence factors’ 365 target cells chosen 424 follicle-stimulating hormone (FSH) 5, 311 vectors used 421, 422, 424–38 biological effects 311, 312, 313–15, 314 non-viral-based systems 421, 422, 432–8 medical uses 320, 321 viral-based systems 421, 422, 424–32 molecular mass 195, 313 gene-based vaccines 444–5 INDEX 471

Genentech (biopharmaceutical company) 6–7, 7 medical uses 319–20 generic biopharmaceuticals 10 recombinant 320–1 genes 41 sales data 319 number in various species 42 sources 319 patenting of 69 veterinary uses 321–3 genetic disease, and gene therapy 438–41 Good Manufacturing Practice (GMP) genetic engineering 37 and contamination by pyrogens 190, 191, 195 vaccine technology affected by 400–2 validation procedures 200, 201 genetically engineered therapeutic proteins 5, 6 Gram-negative bacteria genital warts, treatment of 224, 229 contamination by endotoxin from 190–1 genome(s) 41 lipopolysaccharides on outer membrane 107, 109, gene organization within 42 189, 191, 192 sequencing 59–60 recombinant proteins produced in 8, 105–9 genomics 10, 59–65 granulocyte colony-stimulating factor glass transition temperature (Tg) 169 (G-CSF) 269 glioma-derived growth factor see platelet-derived biological activities 209 growth factor granulocyte macrophage colony-stimulating factor GlucaGen 306 (GM-CSF) 269, 270 305–6 granulocytes 207 initiation of metabolic response 305, 306 growth factors 3, 9, 265–90 glucocerebrosidase 356, 359–60 approved for medical use 266 glucocerebrosides 359 assays for 176 glutamate dehydrogenase 14 wound healing and 279–86 glutamic acid/glutamate 15, 16, 17, 142, 145, 161, (GH) 307–10 162, 357 biological effects 308–9 glutamine 15, 16, 17 mediation by IGFs 281, 309 deamidation of 17, 161, 162 regulation of synthesis and release 307, 308 glutamine amidotransferase 357 therapeutic uses 309–10 glycine 15, 16, 17, 165 growth hormone receptor 307–8 glycoproteins 29–33 approved therapeutic agents 32 haematopoiesis 265, 267, 268 degradation of sugar side chains 161 haematopoietic growth factors 265–79 haematopoietic growth factors 268, 269, 270, 273, colony-stimulating factors 269–72 278, 283 interleukins 268 immunoglobulins 383 haematopoietic stem cells 457 insulin receptor 294 differentiation of 268, 275 interferons 214 in gene therapy 440 interleukin receptors 251 haemoglobin (human) 14 interleukins 242–3 haemoglobinopathies, treatment of 439 microheterogeneity 31, 33 haemophilia A 335–6, 340 oncofoetal antigens 390 haemophilia B 339, 340 role of glycocomponent 31 Haemophilus infl uenzae vaccines 397, 400 sugar side chains 30, 33, 76 haemostasis 329–40 thrombolytic agents 346, 353 mechanisms 329 glycosaminoglycans 313, 315, 341 Hageman factor 330 glycosyltransferases 30 hairy cell leukaemia, treatment of 224, 226 GM-CSF receptor 270, 271 hen eggs, as source of biopharmaceuticals 116 see also granulocyte macrophage colony- heparin 341 stimulating factor hepatitis 228–9 goats, transgenic, milk from 112–15, 345 treatment of 224, 228, 229 gonadotrophins 310–23 hepatitis A vaccines 397, 399, 399 472 INDEX hepatitis B surface antigen (HBsAg) hybridoma technology recombinant 401, 401, 402, 405 interferon production by 225 production of 403, 404 monoclonal antibody production by 198, 374–6 hepatitis B vaccines 397, 400, 401–2, 405 hydrogen bonding, in proteins 27 herpes simplex virus hydrophobic amino acids 17, 146–7, 147 as gene vector 429 hydrophobic interaction chromatography 141, 146–8 thymidine kinase gene 443 hydrophobic interactions, in proteins 27 hexokinase 14 hydroxyapatite chromatography 141, 154–5 high-performance liquid chromatography hyperuricaemia, treatment of 361–2 (HPLC) 155–7, 183–4 advantages 156, 183–4 α-L-iduronidase 356, 362 disadvantages 156–7 IFN-α receptor 215 in downstream processing 155–7 see also interferon-α product purity assessed using 183–4 IFN-γ receptor 215 high-throughput screening 10 see also interferon-γ hirudin 341, 342–4 IL-1β converting enzyme (ICE) 254 binding to thrombin 343 IL-1 receptor 251–2 recombinant 343–4 see also interleukin-1 histidine 15, 16, 17, 144, 145 IL-1 receptor antagonist 7, 252, 253 histidine-tagged proteins 154, 158 IL-2 receptor 243–4 histones 41 binding of monoclonal antibodies 250, 395 homogenization, cell disruption cells capable of expressing 244 by 134–5 see also interleukin-2 hormones 9, 291–328 IL-6 receptor 211 meaning of term 291 IL-11 receptor 211 Humalog (insulin lispro) 298, 301, 302 see also interleukin-11 human anti-mouse antibodies (HAMA) 391 imaging applications, monoclonal antibodies 395 human chorionic gonadotrophin (hCG) 5, 30 immobilized metal affi nity chromatography 153–4 biological effects 311, 312 immune response, agents affecting 59, 242, 246, medical uses 319–20, 322 257 molecular mass 195 immunization 371 human embryonic stem cells, culture of 457–8 see also vaccines human growth hormone (hGH) 307–10 immunoaffi nity chromatography 141, 150, 151 molecular mass 195, 307 applications 140, 150, 337, 339, 340 recombinant preparations 310 immunoassays production of 311 disadvantages 179 therapeutic use 309–10 impurities detected using 185 three-dimensional structure 307 product potency determined by 177–9 human immunodefi ciency virus (HIV) 407 viral assays 198 development into AIDS 408 immunogenicity, proteins 78–9 numbers affected 408 immunoglobulin preparations 372, 373, 374 vaccines 408, 409–10 immunoglobulins 382–3 see also AIDS immunoscintigraphy 384 human (hPTH) 324 coupled with radiolabelled antibodies 390 human serum albumin (HSA) 5, 354–5 immunostimulatory complexes (ISCOMs) 412, 416 as excipient 78, 164 immunosuppressive agents 395 market value 355 IL-2 synthesis prevented by 249 humanized antibodies 392–3, 394 immunosurgery 457 examples (listed) 380, 381 immunosurveillance 379 Humaspect 381, 392 impurities, medical consequences 174 Humulin 7, 8, 106, 298 in vitro fertilization procedures 457 INDEX 473 inclusion bodies (in heterologous proteins) 107, 158 production of 225–7 eliminating/reducing 108–9 receptors 215 Increlex 266 side effects 227, 230, 235, 235 Indimacis-125 380, 390 toxicity 234–5 Infergen 224, 227, 228 interferon-α (IFN-α) 226–9 infl ammatory response, agents affecting 59, 252, biochemistry 213–14 257–8 medical uses 224, 226, 228, 229 inhalable insulin product 11, 71, 298, 304 molecular mass 195, 213 inhibin(s) 311, 313, 315, 317–19 production of 226–7, 227, 228 initial product recovery 134 interferon-β (IFN-β) 8, 229–31 insect cell based systems, as source of mechanism of action 231 biopharmaceuticals 118–20 medical uses 229–31 insulin 291–305 molecular mass 195, 214 alternative administration routes 304–5 production of 230, 231 animal-derived 294, 296 interferon-γ (IFN-γ) 214, 232–4 availability 296 biological activities 209, 219–20, 266 immunogenicity 296 medical uses 232–4 engineered 298, 301–4 molecular mass 195, 214 formulation of products 297, 300 interferon-ω (IFN-ω) 120, 236 infusion pump systems 304–5 interferon-tau (IFN-τ) 236, 236–7 inhalable product 11, 71, 298, 304 interferon JAK–STAT pathway 218–19 metabolic effects 292 interferon 214 molecular mass 195, 293 interferon-stimulated response elements molecular structure 293–4 (ISREs) 214, 218 production of interleukin(s) 3, 5, 9, 241–54 animal-derived products 294, 296 biological activities 242, 266 by recombinant DNA technology 297, 299, 301 cells capable of producing 241, 242 recombinant human 7, 8, 106, 297, 298 molecular mass 195 structure 14 interleukin-1 (IL-1) 190, 195, 251–4 Swiss-Prot database entry 21, 22 biological activities 209, 252 Insulin Aspart 298, 301 biotechnology 253–4 insulin detemir 298, 303 cells capable of producing 242, 251, 252 insulin glargine 298, 301, 303 molecular mass 195, 251 insulin glulisine 298 receptors 251–2 insulin lispro 298, 301, 302 three-dimensional structure 251 manufacture of 302 interleukin-2 (IL-2) 14, 242–50 three-dimensional structure 303 and cancer treatment 246–8 insulin receptor 294, 295 in fused proteins 250–1 insulin-dependent diabetes mellitus (IDDM) 292 high-affi nity receptor 243–4 insulin-like growth factors (IGFs) 280–2 and infectious diseases 248–9 biological effects 266, 281–2 inhibition of activity 249–50 molecular mass 195 molecular mass 195, 243 Insuman 298 production of 246 interferon(s) 3, 5, 9, 212–37 receptors 243–4 antiviral activities 219 safety issues 249 enzymatic mechanisms 220–3 three-dimensional structure 243 assays for 176 interleukin-11 (IL-11) 254 biological activities 212 interleukin-12 (IL-12) 195 biotechnology 224–36 interleukins, as haematopoietic growth factors 268 classifi cation 212–13 International Conference on Harmonization molecular mass 195 (ICH) 101 474 INDEX intracellular immunization 444 ligands Intron A 7, 8, 224, 226, 226–7 affi nity chromatography 148, 149 medical conditions for which use approved 226 dyes as 152, 153 investigational new drug (IND) application 92–3 Limulus amoebocyte lysate (LAL) test 193–4 ion-exchange chromatography advantages compared with rabbit test 193 analysis by 184 disadvantage 194 DNA removal by 196 validation studies 194 downstream processing by 140, 141, 142, 144–6 lipase(s) 365 endotoxin removal by 195 lipoplexes, in gene delivery systems 434 Iplex 266 lipopolysaccharide (LPS), on Gram-negative isoelectric focusing 181–2 bacteria 107, 109, 189, 191, 192 isoelectronic point (pI) 144 liposomes 412, 415–16 in chromatofocusing 155 Liprolog (insulin lispro) 298 in isoelectric focusing 181 Listeria monocytogenes 249 Lowry protein assay method 179 JAK–STAT pathway 215–18 (LH) 76, 311 interferon-induced 218–19 biological effects 311, 312, 315 proteins inhibiting 218, 219 molecular mass 195, 315 signal transduction process mediated lymphocyte-activating factor (LAF) see interleukin-1 by 216, 217 lymphocytes 206–7 Janus kinases (JAKs) 216 lymphokine-activated killer (LAK) cells 245, 248 see also JAK–STAT pathway lymphotoxin see tumour necrosis factor β Japan, drug registration in 100 lyophilization see freeze drying lysine 15, 16, 17, 144, 145, 165 Kepivance 7, 106, 266, 285 lysine affi nity chromatography 133, 140 keratinocyte growth factor 7, 106, 266, 284, 285 lysozyme 14 Kineret 7, 253–4 kininogen 331 macrophage colony-stimulating factor (M- knockout animal studies 61 CSF) 269–70 macrophages 206 labelling of product 169, 171 Macugen 8, 10, 419, 453, 454 lac operon 43 major histocompatibility complex (MHC) antigens, lactase 365, 366 stimulation of 220 lactose, hydrolysis of 366 malaria 248 lactose intolerance 366 male reproductive cycle, hormones in 318 Lantus (insulin glargine) 298, 301, 303 Maloney murine leukaemia virus (MoMuLV) 426 laronidase 356, 362, 363 mammalian cell culture systems 127–9

LD50 studies 82 bioreactor design 126, 128 lectin affi nity chromatography 150–2 microcarriers 128–9 limitations 151–2 manufacturing licence 57 lectins (listed) 151 marketing approval 57 leech-derive anticoagulant 342 mass spectrometry 184–5 Legionella pneumophila 249 Maxam–Gilbert chemical sequencing method 45 leishmaniasis, treatment of 233 measles vaccines 398, 399 leukaemia, treatment of 226, 356 266 leukaemia inhibitory factor (LIF) 266 266 Leukine 266 megakaryocytes 279 leukocytes 205, 206–7 meningococcal vaccines 398, 400 classifi cation 207 menotrophin(s) 319 Levemir (insulin detemir) 298, 303 medical uses 319 Leydig cells 315, 318 Merispace 118 INDEX 475 messenger RNA (mRNA) 38 neurodegenerative diseases, treatment of 282, 286 blocking of translation step 447 3, 211, 286 editing in eukaryotes 43, 44, 63 cells targeted by 266, 286 metal chelate affi nity chromatography 153–4 medical applications 267, 286 applications 120, 154, 158 neutropenia 271 Metalyse 346, 349 causes 271 methionine 15, 16, 17 treatment of 266, 271, 272 oxidation of 17, 162, 163 neutrophils 207 microbial contaminants 196–7 new drug application (NDA) 92, 94–5 microbial fermentation 124–7 contents 94 non-pharmaceutical products using 125 review process 94, 95 plasmid DNA manufacture using 436 NK cells, activation by interleukins 245, 247 microfi ltration 134, 136 nonacog alfa 339 milk, transgenic animals, as source of therapeutic Novolin 298 proteins 10, 112–15, 345, 354 Novolog (Insulin Aspart) 298 mineral-based adjuvants 412, 413–14 NovoRapid (Insulin Aspart) 298 molecular biology 37, 38 NovoSeven 335, 340 molecular mass nuclear magnetic resonance (NMR) analysis, biopharmaceuticals (listed) 195 structural determination of proteins 65 proteins (listed) 14 nucleases 70, 136, 356, 357–8 monoclonal antibodies 374–95 nucleic-acid-based therapeutic agents 8, 9, 419 binding to IL-2 receptor 250, 395 nucleic acid sequence databases 21 imaging applications 395 nucleic acid sequencing 45, 46 production of 198, 374–6 nucleic acids radiolabelled 383–4 bases in 38, 39 therapeutic applications 378–9, 395 function 38 transplantation-related 395 purifi cation of 43–5 monoclonal antibody based products 8, 9, 380–1 removal in downstream processing 136 monoclonal antibody technology 5 structure 38–41 mononuclear phagocytes 206 see also DNA; RNA multiple drug resistance (MDR-1) gene product 444 nucleotides 38, 39 (MS), treatment of 229–31 null cells 207 mumps vaccines 398, 399, 399 see also NK cells muramyl dipeptide (MDP), as adjuvant 412, 414–15 Nutropin 7, 310 murine monoclonals, antigenicity 391–2 mutagenicity tests 83, 84, 85 β-oestradiol 316, 317 mycobacterial infections 234, 248 oestriol 316, 317 mycobacterial species, as adjuvants 412, 414 oestrogens 314, 316 Mylotarg 381, 385–6 biological actions 314, 317 Myozyme, preclinical tests 84, 85 oestrone 316, 317 oil-based emulsion adjuvants 412, 414 N-terminal sequencing, protein characterization by 188 2Ј-5Ј oligoadenylate synthestase (2,5- nasal delivery route 73–4 Ansynthetase) 220, 221 naturally sourced proteins 5 oligonucleotides generated by 220, 222 Neorecormon 266, 276 oligos see antisense oligonucleotides production of 277 OM-1 implant 95 nerve growth factor (NGF) 195, 286 oncofoetal antigens 390 Nespo 266, 276 Ontak 6, 250–1 Neulasta 7, 266 Optisulin (insulin glargine) 298, 301, 303 Neumega 254 oral delivery systems 70–1 Neupogen 7, 266, 272 limitations 71 476 INDEX oral mucositis, treatment of 7, 106, 266, 285 plasminogen 346, 347 Orthoclone OKT-3 380, 395 plasminogen activation 350, 351–2 osteoporosis, treatment of 324 platelet-derived growth factor (PDGF) 70, 283–4 osteosarcoma-derived growth factor see platelet- cells targeted by 266, 283 derived growth factor platelets 279 ovarian follicle pluripoietin 269 granulosa cells 313, 314, 316, 318 pluripoietin-α 270 theca cells 314, 315, 316, 318 pluripotential stem cells 265 Ovitrelle/Ovidrel 321, 322 pneumococcal vaccines 398, 400 poliomyelitis vaccines 398, 399, 399 ‘packing cells’, in retroviral vector 426 polycistronic mRNA 43 packing of products 171 polyclonal antibody preparations 371–3, 374 Paget’s disease 324–5 poly(ethylene glycol) (PEG) interactions 266 interferons 227–8 pancreatin 365 synthetic gene delivery systems 434 papain 356, 364 polyethylenimine, nucleic acids precipitated by 136 paracellular transport 72, 73 polylysine 433 patent, meaning of term 68 in gene delivery systems 434 patentability criteria 68 polymerase chain reaction (PCR) technique, DNA patenting 67–70 cloning via 51–3 in biotechnology 68–70 polynucleotides, structure 38, 40 Pegasys 224 polyols, as stabilizing agents 165 PegIntron A 224, 227 polypeptide growth factors 265, 266 peptide bond(s) 18–19 polypeptides 13, 15 hydrolysis of 160 amino acid sequence determination 19–21 see also polypeptides bonding in 18–19 peptide mapping, protein characterization by 186–8 characterization of 19–21 peptide vaccines 402–3 disulfi de exchange in 162, 163 phage display technology 376–8 factors affecting conformation 16 phagocytes structure-prediction methods 28–9

expression of IgG Fc receptors on 233, 234 synthesis 22–3 interferon-γ as activator 233 polysorbates, as excipients 78, 166 reactive oxygen species produced by 233 Porcilis pesti (vaccine) 9, 411–12 pharmaceutical industry, history 2–3 post-marketing surveillance studies 58, 86–7 pharmacodynamics 79–80 potency testing 173, 175–9 pharmacogenetics 65–6 poxviruses, as vaccine vectors 403, 405 pharmacokinetics 74–80 preclinical studies/trials 57, 74, 83–4 pharmacology 74 tests undertaken during 75, 83–4, 85 phase I trials 86 pregnant mare serum gonadotrophin (PMSG) 311, phase II trials 86 315, 323 phase III trials 86 prekallikrein 331 phase IV trials 86–7 proaccelerin 330 phylogenetic profi ling 61 Procrit 8, 266 plasma proteins, in human blood (listed) 354 product analysis 173–203 plasma thromboplastin antecedent 330 product homogeneity, determination of 181–2 plasmid DNA product potency 175–9 as gene vector 432–6 product purity 175 manufacture of 436–8 production see downstream processing; upstream impurities 436, 437 processing purifi cation of 437–8 proinsulin, proteolytic processing of 293 plasmids 41, 47–8 prokaryotes, genes 43 plasmin 346, 347 Proleukin 246 INDEX 477 prosthetic groups (in proteins) 14 separation methods 63–4 protamine–zinc–insulin suspensions 300, 301 stability and folding 27–9 proteases 160, 364, 365 β-strands/sheets 25 inhibitors 161 structural characterization of 66–7 protein A chromatography 150 structure 15–18, 23–6 protein-based contaminants 173–4 sulfation of 30, 34 clinical signifi cance 174 tertiary structure 26 detection of 180–9 analysis 188 electrophoretic methods 180–2 structural characterization of 67 high-performance liquid chromatography 183–4 three-dimensional structure 26 immunological approaches 185–9 see also glycoproteins; polypeptides mass spectrometry 184–5 proteolytic enzymes 364 protein C 341 proteomics 10, 62–4 activated 345 prothrombin 330, 332, 333 protein engineering 53–4 pUC18 plasmid 48, 49 protein– interactions, factors affecting 149 pulmonary delivery 71–2 protein sequence databases 21 factors affecting bioavailability 72 protein sequencing 19, 188, 190 physiological barriers 72 protein structure databanks 21, 65 Pulmozyme 359 protein(s) purifi ed water 131 amidation of 30, 34 generation of 134 amino acids in 15–17, 142, 144, 145 purine metabolism 361 sequence determination 19, 21, 188, 190 pyrogens 189–90 assay methods 179–80 detection of 191–5 γ-carboxylation of 30, 33, 34 deamidation of 161, 162 quality control (QC) procedures 166, 169, 171, 173 denaturation of 13, 159 determination of concentration 179–80 rabbit pyrogen test 191–3 domains 26 disadvantages 192–3 examples 14 radioimmunoassays (RIA) 177 factors affecting biological activity 159–63 radiolabelled antibodies 383–4 glycosylation of 29–33 coupled with immunoscintigraphy 390 α-helix 23–5 Ramachandran plot 19 β-hydroxylation of 30, 33, 34 randomized double-blind trial(s) 88, 89 imide formation 161 Rapilysin 346, 348 immunogenicity 77, 78–9 Rebetron 224 isoelectronic point 144 Rebif 224, 230 loop structures 25 receptor-mediated endocytosis 76, 77, 435, 450 metabolism/excretion of 76 recombinant blood factor preparations 5 modes of action 79–80, 81 recombinant DNA (rDNA) technology 4–5, 37, 46 modifi cation of pharmacokinetic profi le 77, 79, 80 advantages 5 N-linked glycosylation of 30–1, 32 blood factors 337, 339, 340 O-linked glycosylation of 30, 31, 32 erythropoietin 276 pharmacokinetics 75–7 gonadotrophins 320–1 post-translational modifi cations (PTMs) 29–34 insulin 297, 299 primary structure 15–18 interferons 225 structural characterization of 67 interleukins 246 proteolytic degradation of 160–1 monoclonal antibodies 392 recombinant production of 46–7 vaccines 400–2 secondary structure 23–5 recombinant gonadotrophins 320–1 analysis 188 recombinant human insulin 7, 8, 106, 297, 298 structural characterization of 67 recombinant proteins, purifi cation of 157–8 478 INDEX

Recombinax vaccine 401, 403 severe combined immunodefi ciency (SCID) 440 Refl udan 343 X-linked 428 production of 343–4 sheep, transgenic, milk from 112–15, 354 Regranex 266, 280, 284 short interfering RNAs (siRNAs) 452 regulation of biopharmaceutical products 57–8 signal transducers and activators of transcription regulatory authorities 89–101 (STATs) 216 in Europe 96–100 functional domains 217 in Japan 100 ligands activating 218 in USA 90–5 see also JAK–STAT pathway Remicade 8 signal transduction mechanisms renal function, effect of IGFs 282 cytokine receptors 211 Replagal 360 GM-CSF receptor 270, 271 reproductive toxicity studies 82, 85 insulin receptor 294, 295 restriction endonucleases (REs) 47 interferons 214, 218 commercially available 49 interleukins 241–2, 252 Retavase 346, 348 JAK–STAT pathway 216, 217 reteplase 348 tumour necrosis factor 259–60 retroviruses silkworm (Bombyx mori), vaccines produced in 118, cancer induced by 389–90 120 as gene vectors 424–8 silver-binding protein assay method 179, 180 life cycle 424–5 simple proteins 13 Revasc 344 Simulect 380, 395 reverse-phase chromatography 148 sindbis virus reverse-phase HPLC 184 as gene vector 430–1 in insulin production 297, 299 generation of 430 ribose 38, 39 single nucleotide polymorphisms (SNPs) 65–6 see also RNA identifi cation and characterization of 66 ribosomal RNA (rRNA) 38 site-directed mutagenesis 5, 53 ribozymes, mRNA cleavage using 452–3 size-exclusion chromatography 142 Rituxan 7, 8 see also gel-fi ltration chromatography RNA 38 size-exclusion HPLC 184 RNA interference, gene silencing by 451–2 see insulin-like growth factors Roferon A 224 somatotrophin see human growth hormone roller bottle systems 128 sources of biopharmaceuticals 105–20 Rosetta stone approach 61 animal cell culture systems 109–10 rubella vaccines 398, 399 E. coli 105–9 fungi 111 S-oligos 449 insect cell based systems 118–20 Saccharomyces cerevisiae, biopharmaceuticals transgenic animals 111–16 produced in 8, 110, 111 transgenic plants 116–18 safety testing 173 yeast 110–11 salt bridges, in proteins 27 Staphylococcus aureus, protein produced by 150 Sanger–Coulson enzymatic sequencing method 45 staphylokinase 346, 351–3 saponins 412, 415 STATs see signal transducers and activators of 266 transcription SDS polyacrylamide gel electrophoresis (SDS- stem cells 11, 265, 457–60 PAGE) 180–1 in gene therapy 440 seminal fl uid, as source of therapeutic proteins 115 see also adult...; embryonic...; haematopoietic stem studies 61 cells serine 15, 16, 17 sterilizing fi ltration 166, 167–8, 190, 197 Sertoli cells 313, 318 streptokinase 346, 350 INDEX 479 structural genomics 64–5 tissue factor 330 Stuart factor 330 tissue plasminogen activator (tPA) 9, 346–8 subtilisin 14 domains constituting 26, 346 subunit vaccines 401–2 fi rst-generation 348 sulfa drugs 3, 4 modifi ed/engineered 6, 346, 348–9 superoxide dismutase (SOD) 356, 363 purifi cation of 133, 140, 348, 349 surface antigens tissue-based therapies 453–60 vaccines 400 TNKase 346 see also tumour surface antigens toxicity studies 80, 82–4 surfactants, as stabilizing agents 165–6 see also clinical trials survivin 448 toxicity testing, fi nished products 199 swine-fever-based vaccines 9, 119 toxin-conjugated monoclonal antibodies 384–6 Swiss-Prot (protein) database 21, 21, 22 transcytosis 72, 73 Syntex adjuvant 415 transdermal delivery systems 74 synthetic vaccines 402–3 transfer RNA (tRNA) 38 systematic evolution of ligands by exponential transforming growth factors (TGFs) 284–6 enrichment (SELEX) 453 cells targeted by 266, 285–6 medical applications 267, 286 T-cell growth factor see interleukin-2 transgenic animals T-lymphocytes 207 generation of 111–12 activation by interleukins 245, 252 mammary-specifi c expression of proteins 112–15, tagged proteins, purifi cation of 154, 158 345, 354 tasonermin 261 patenting of 69 technetium-labelled monoclonal as source of biopharmaceuticals 10, 111–16 antibodies 384 time lag to fi rst production batch 114–15, 114 tenecteplase 346, 349 typical milk yields 114 teratogenicity studies 82 transgenic plants 315, 316 advantages and disadvantages 117 tetanus vaccines 398, 400 patenting of 69 thioredoxin 108 as source of biopharmaceuticals 116–18 threading technique (in structure-prediction of transmucosal delivery systems 74 polypeptides) 29 transplantation 454 threonine 15, 16, 17, 165 and monoclonal antibodies 395 threonyl-MDP, as adjuvant 415 see also whole blood transfusion thrombin 330, 332, 333, 334 trehalose dimycolate (TDM), as adjuvant 412, 414 binding to hirudin 343 triazine dyes 152, 153 biological activities 332, 333, 334, 342 Tritanrix 401, 402 thrombin inhibitors 342–5 trophoblastin 235–6 thrombocytopenia, treatment of 278–9 trypsin 356, 364 thrombolytic agents 9, 345–55 tuberculosis vaccine 397, 398 thromboplastin 330 tumour cells, ‘sensitivity’ gene introduced thrombopoiesis, stimulation of 254 into 443 (TPO) 278–9 tumour detection/destruction cells targeted by 266, 267 antibody-based strategies 383–6 medical applications 267 gene therapy approaches 442–3 molecular mass 195, 278 tumour immunology 379, 382–9 thrombosis 345 antibody-mediated approach 383–6, 388–9 thrombus formation 340 drug-based 386–7 Thyrogen 324 tumour necrosis factor (TNF) 9, 14, 255–61 -stimulating hormone (TSH) 323–4 symptoms mediated by 261 thyrotrophin alfa 324 therapeutic aspects 260–1 480 INDEX tumour necrosis factor α (TNF-α) 14, 59 vaccinia virus, as vector 405, 406 biochemistry 255–6 validation studies 199–202 biological activities 256–7 glossary of terms 200 cellular sources 255 for LAL assay 194 immunity affected by 257 vascular endothelial growth factor (VEGF), inhibition infl ammatory response affected by 257–8 of action 394, 454 molecular mass 195 veterinary biopharmaceuticals 9, 321–3 physiological inducers 256 veterinary vaccines 9, 411–12 three-dimensional structure 255, 256 Vibragen Omega 9, 120, 224 tumour necrosis factor receptors 258–60 Viraferon 224 tumour suppressor-based gene therapy 442, 443 ViraferonPeg 224, 227 tumour surface antigens (TSAs) 379 viral assays 198 complications caused by 390–1 viral clearance (from product) 197 tumour-associated antigens 389–91, 410 validation of 201 tumour-infi ltrating lymphocytes (TILs), genes viral contaminants 197 introduced into 442 viral hepatitis, treatment of 224, 228, 229 Twinrix 401, 402 viral vectors (in gene therapy) 421, 422, 424–32 two-dimensional electrophoresis 181 adeno-associated 429 adenoviral 428–9 ulcers 279–80 herpes-based 429 ultrafi ltration 137–9, 197 manufacture of 431–2 cartridge-type fi lters 138 retroviral 424–8 hollow-fi bre membranes 139 advantages 426 membrane materials 137 disadvantages 427–8 minimization of concentration polarization 137 sindbis 430–1 unique surface antigen (USA) 379 Virtron 224 upstream processing 120–9 vitamin K antimetabolites 341 urate oxidase 356, 361, 362–3 Vitravene 8, 419, 450 uric acid 361–2 von Willebrand’s disease 336 oxidation of 361, 362 von Willebrand’s factor 335, 336 urine, transgenic animals, as source of therapeutic proteins 5, 115 warfarin 341 urokinase 346, 350–1 water for injections (WFI) 131 US patent law 68–9 generation of 134 USA, drug registration in 90–5 white blood cells see leukocytes whole blood transfusion 455–6 vaccine vectors 403, 405–7 whole-cell-based therapeutic products 11 vaccines 9, 396–412 ‘whooping cough’ vaccine 403 adjuvants for 412–16 wound healing process, effect of growth antigen-based 400 factors 279–86 attenuated/dead/inactivated bacterial 398–9 attenuated/inactivated viral 399 X-ray crystallography, structural determination of classifi cation 397 proteins 65 egg-based production of 116, 399 xanthine, oxidation of 361 gene-based 444–5 Xigiris 345 global market value 396 impact of genetic engineering 400–2 yeasts, as source of biopharmaceuticals 110–11 toxoid-based 399–400 yellow fever vaccines 398, 399 traditional 396–400 transgenic plant-based production of 117–18 Zenapax 380, 395 veterinary 9, 411–12 zinc–insulin suspensions 300, 301