Manufacturing of Gene Therapeutics Methods, Processing, Regulation, and Validation Edited by G Subramanian Littlebourne, Kent, England Kluwer Academic / Plenum Publishers New York, Boston, Dordrecht, London, Moscow Library of Congress Cataloging-in-Publication Data Manufacturing of gene therapeutics: methods, processing, regulation, and validation/ edited by G Subramanian p cm Includes bibliographical references and index ISBN 0-306-46680-5 Gene therapy Genetic vectors Genetic transformation I Subramanian, G., 1935[DNLM: Gene Therapy Drug Approval Genetic Engineering Genetic Vectors—biosynthesis Pharmaceutical Preparations QZ 50 M294 2001] RB155.8 M36 2001 616/042—dc21 2001038587 ISBN: 0-306-46680-5 ©2002 Kluwer Academic / Plenum Publishers, New York 233 Spring Street, New York, N.Y 10013 http://www.wkap.nl/ 10 A C.I.P record for this book is available from the Library of Congress All rights reserved No part of this book may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, microfilming, recording, or otherwise, without written permission from the Publisher Printed in the United States of America Preface Advances in molecular biology and recombinant DNA technology have accelerated progress in many fields of life science research including gene therapy A large number of genetic engineering approaches and methods are readily available for gene cloning and therapeutic vector construction Significant progress is being made in genomic, DNA sequencing, gene expression, gene delivery and cloning Thus gene therapy has already shown that it holds great promise for the treatment of many diseases and disorders In general it involves the delivery of recombinant genes or transgenes into somatic cells to replace proteins with a genetic defect or to transfer with the pathological process of an illness The viral and non-viral delivery systems may hold the potential for future non-invasive, cost effective oral therapy of genetically based disorders Recent years have seen considerable progress in the discovery and early clinical development of a variety of gene therapeutic products The availability, validation and implementation have enabled success but also for testing and evaluation New challenges will need to be overcome to ensure that products will also be successful in later clinical development and ultimately for marketing authorisation These new challenges will include improvements in delivery systems, better control of in-vivo targeting, increased level transduction and duration of expression of the gene, and manufacturing process efficiencies that enable reduction in production costs Perhaps profound understanding of regulated gene design may result in innovative bioproducts exhibiting safety and efficacy profiles that are significantly superior to those achieved by the use of naturally occurring genes This procedure may considerably contribute to fulfil standards set by regulatory authorities This book aims to project an overview of the current advances in the field of gene therapy and the methods that are being successfully applied in the manufacture of gene therapeutic products I am indebted to the international group of contributors who have shared their practical knowledge and experience Each chapter represents an overview of its chosen topic Chapters one and two provide an overview of gene therapy and gene therapy for cancer Gene self-assembly and gene expression are discussed in chapters three and four Genotype and response to cytotoxic gene therapy is reviewed in chapter five Vector assembly and gene transfer is discussed in chapters six and seven Plasmid manufacturing is reviewed in chapter eight The importance of quality control and assurances and the analytical methods are discussed in chapters nine and ten Chapter eleven provides an insight into validation aspects in gene therapy and gene delivery is reviewed in chapter twelve The importance of regulatory issues and guidelines are reviewed for the American market in chapter thirteen and the European market in chapter fourteen, and chapter fifteen discusses the regulatory compliance in contract manufacturing environment Finally, chapter sixteen discusses the risk assessment in gene therapy My thanks to the contributors for the extensive diligence and their patience and goodwill during the production of the book; they deserve the full credit for the source of the book It is hoped that this book will be of great value to all those who are engaged in the field of gene therapy and that it will stimulate further progress and advancement in this field to meet the ever increasing demands I should be most grateful for any suggestion, which could serve to improve future editions of this book My deep appreciation to Jo Lawrence of Kluwer Academic/Plenum Publishers for her continuous patience, encouragement and help in guiding all of us through the preparation and the completion of this book G Subramanian Contributors Akshay Anand Department of Immunopathology Post Graduate Institute of Medical Education and Research Chandigarh, India Sunil K Arora Department of Immunopathology Post Graduate Institute of Medical Education and Research Chandigarh, India Joy A Cavagnaro President, Access Bio LC Leesburg VA 20177-1400, USA Nancy Chew President, Regulatory Affairs, North America LLC P.O.BOX 72375 Durham NC 2772, USA Odile Cohen-Haguenauer Laboratorie TGOM & Service d'Oncologie Medicale Hopital Saint-Louis 1, avenue Claude Vellefaux 75475 Paris CEDEX 10, France Peter Daniel Department of Haematology, Oncology and Tumour Immunology University Medical Centre Charite Campus Berlin-Buch Humboldt University of Berlin Lindenberger Weg 13125 Berlin, Germany Linh Do Berlex Biosciences 15049 San Pablo Avenue P.O.BOX 4099 Richmond CA 94804-4089, USA Vladimir I Evtushenko Laboratory of Genetic Engineering Research Institute of Roentgenology and Radiology St Petersburg 189646, Russia James G Files Berlex Biosciences 15049 San Pablo Avenue P.O.Box 4099 Richmond CA 94804-4089, USA Erwin Flaschel PlasmidFactory GmbH & Co KG Meisenstrasse 96 D-33607 Bielefeld, Germany Karl Friehs University of Bielefeld Postfachl00131 D-33501 Bielefeld, Germany Bernhard Gillissen Department of Haematology, Oncology and Tumour Immunology University Medical Centre Charite Campus Berlin-Buch Humboldt University of Berlin Lindenberger Weg D-13125 Berlin, Germany Clague P Hodgson Nature Technology Corporation 4701 Innovation Drive Lincoln Nebraska 68521,USA John Irving Berlex Biosciences 15049 San Pablo Avenue P.O.Box 4099 Richmond CA 94804-4089, USA Juan Irwin Berlex Biosciences 15049 San Pablo Avenue P.O.Box 4099 Richmond CA 94804-4089, USA John Jenco Dow Biopharmaceutical Contract Manufacturing Services 50 East Loop Road Stony Brook NY 11790, USA Jaspreet Kaur Department of Biochemistry Postgraduate Institute of Medical Education and Research Chandigarh, India Steven S Kuwahara Kuwahara Consulting PMB #506 1669-2 Hollenbeck Avenue Sunnyvale CA 94087-5042, USA Mark Lawler Department of Haematology and Oncology St Patrick Dun Research Labs James's Street Dublin 8, Ireland Elisabeth Lehmberg Berlex Biosciences 15049 San Pablo Avenue P.O.Box 4099 Richmond CA 94804-4089, USA Bruce Mann Berlex Biosciences 15049 San Pablo Avenue P.O.Box 4099 Richmond CA 94804-4089, USA Michael T McCaman Berlex Biosciences 15049 San Pablo Avenue P.O.Box 4099 Richmond CA 94804-4089, USA Stephen Morris BioReliance 14920 Broschart Road Rockville MD 20850-3349, USA Munishi Mukesh National Bureau of Animal Genetics Resources (ICAR) Karnal, India Peter K Murakami Berlex Biosciences 15049 San Pablo Avenue P.O.BOX 4099 Richmond CA 94804-4089, USA Chris Murphy BioReliance 14920 Broschart Road Rockville MD 20850-3349, USA Jeffrey W Nelson Berlex Biosciences 15049 San Pablo Avenue P.O.Box4099 Richmond CA 94804-4089, USA Eirik Nestaas Berlex Biosciences 15049 San Pablo Avenue P.O.BOX 4099 Richmond CA 94804-4089, USA Erno Pungor, Jr Berlex Biosciences 15049 San Pablo Avenue P.O.Box 4099 Richmond CA 94804-4089, USA Martin Schleef PlasmidFactory GmbH & Co KG Meisenstrasse 96 D-33607 Bielefeld, Germany Torsten Schmidt PlasmidFactory GmbH & Co KG Meisenstrasse 96 D-33607 Bielefeld, Germany Gail Sofer BioReliance 14920 Broschart Road Rockville MD 20850-3349, USA Isrid Sturm Department of Haematology, Oncology and Tumour Immunology University Medical Centre Charite Campus Berlin-Buch Humboldt University of Berlin Lindenberger Weg D-13125 Berlin, Germany Mei P Tan Berlex Biosciences 15049 San Pablo Avenue P.O.Box 4099 Richmond CA 94804-4089, USA Joseph A Trai-Na Berlex Biosciences 15049 San Pablo Avenue P.O.Box 4099 Richmond CA 94804-4089, USA Spencer Tse Berlex Biosciences 15049 San Pablo Avenue P.O.BOX 4099 Richmond CA 94804-4089, USA Dominic K Vacante BioReliance 14920 Broschart Road Rockville MD 20850-3349, USA 345 Index terms Links Diseases (Continued) Wilson's documentation 61 324 L-DOPA 10 Dopamine 10 DNA fragments 109 removal 239 E Edman's degradation 211 ELISA 207 208 221 Electron microscopy 202 micrograph 179 Electrophoresis agarose gel 156 Capillary gel 156 Electroporation 140 142 EMEA 295 Endocytosis 259 Endosomal membrane 136 Endosomytic peptides 148 Endonucleases Endotoxin 36 69 108 181 190 234 320 Enzymatic digestion 239 Enzyme stabilisers 103 This page has been reformatted by Knovel to provide easier navigation 346 Index terms EPG motifs Epipodophyllotoxins Links 157 25 Ethical reviews 293 Ethidium bromide 207 Exons 49 Expressions analysis 108 systems 278 transgenic 209 210 viral gene 209 210 F Facility design 282 FBS 233 FDA 12 293 295 Fermentation 157 315 Fibroblast 279 Flame Photometry 230 Flavoprodol 76 Flow cytometry 184 Fluorescence 184 Fluorocytosine 86 Formulation 313 Fractionation 313 Freeze thaw 236 This page has been reformatted by Knovel to provide easier navigation 347 Index terms Links G Gancyclovin Gel electrophoresis 21 245 161 319 Gene amplification 33 cloning 99 101 construction 33 cytokine 18 delivery 245 disorder 240 disease 245 expression 45 function 246 inserted manipulation 109 marker 278 marking material molecular muted 274 mutation regulation 45 retinoblastoma 62 self assembly 253 stability synthese therapeutics 53 33 59 186 109 60 transfer 135 252 transcription 258 This page has been reformatted by Knovel to provide easier navigation 348 Index terms Links Gene (Continued) tumour suppressor vaccination 60 Gene therapy ex vivo 194 germ-line infusion in vivo protocol 161 somatic Genome adenovirus 236 manipulation 106 viral 206 Genomic DNA 163 Genetic identity molecular mutation Genotoxic 61 Germ-line therapy 331 336 GLP 291 Glycolysis Glycosylation Glucocerebrosidase 75 211 205 GMO 290 GMP compliance 173 GMP guidelines 155 Godd clinical practice 173 This page has been reformatted by Knovel to provide easier navigation 349 Index terms GPAT Links 20 Gramicidin-S 250 Granulocyte macrophage 18 Glutathione S transferase 25 GSF GTAC 23 GVHD 23 H Haemoglobinopathies Haemophilia 13 HEK 293 Haemoagglutination 187 Herpes simplex thymidine kinase Hepatocellular carcinoma Hepatocytes 60 254 Hexamine cobalt 103 Histone octamer 46 HIV 61 Hollow fibre ultrafiltration Hormone receptors 289 237 46 Host cell protein 241 234 Host chromosome DNA 163 Hypercholesteremia 247 Hyperthermia 70 Hyperphosphorylation 65 Hypoxia 75 This page has been reformatted by Knovel to provide easier navigation 350 Index terms Links I IBC 274 293 Icosahedral nucleocapsid 257 Identity 319 Immunoassay 208 251 Immunodeficiency 247 273 289 Immunofluorescence 254 Immunogenic sequences 156 Immunogenicity 77 202 Immunoprecipitation 73 IND 274 293 Infectivity 209 242 Interferon 19 Interleukin 36 Introns 49 IOM 275 IRB 274 293 Irradiation 74 Issues compliances 282 efficiency 281 global 284 regulatory 282 safety 279 K Keratinocytes Kinase inhibitor 74 This page has been reformatted by Knovel to provide easier navigation 76 351 Index terms Kinetic PCR Links 163 L LDL Lentiviral 106 Leukaemia Leukophoresis Ligands 248 Lipofection 262 Liposomes 79 Liposomal gene delivery 262 Lipopolysaccharides 137 163 Lysis Cell 316 chemical 316 enzymatic 316 mechanical 317 Lysozomes 316 M MALDI TDF MS 205 214 216 Mass spectroscopy 205 MCA 284 MDR 25 Melanoma 18 Mesenchymal-stem cells Mesothelioma Metabolic disorders 296 86 This page has been reformatted by Knovel to provide easier navigation 352 Index terms Links Microarray 102 Microcarrier 233 279 Microfluidiser 234 236 Microinjection 145 Microorganism 234 Microprojectiles 144 Microsomal assays 333 Mitochondria 73 Mitochondrial factors 69 membrane 73 Mitosis 61 Mutogenicity 333 Mutational analysis 135 Mycloblast 279 Mycoplasma 173 N Naked DNA 156 NIH 275 276 Non Hodgkin's Lymphoma 27 Non viral system 137 Nuclear targeting 260 Nucleases 185 Nucleosome 46 Nystagmus 330 This page has been reformatted by Knovel to provide easier navigation 77 353 Index terms Links O OBA 274 Occupational safety 275 Oligodeoxynucleotides Oncogene 33 Oncoprotein 75 Ovarian cancer 77 P Palindromes 34 Pancreatic adeninocarcinoma 79 Pancreatic Dnase 113 Particle determination 242 Pathogenicity 332 PCR 177 183 187 208 315 316 319 333 PERCO Peptidyl transferase Pharmacopea Phosphorylation Pharmacokinetics Phorbal esters 213 51 228 63 205 109 53 Picogreen 120 207 212 PLA 296 Plasmid 311 DNA PNP 106 155 264 This page has been reformatted by Knovel to provide easier navigation 354 Index terms Links Polyethylenimine 269 Polygenic disorders 174 Polysine 259 Polyvinylpyrrolidine 103 Porcine cells 174 Potency 321 242 Primary cells 147 recovery 310 Procaspase 72 Process, control system 241 development 313 residuals 321 validation 181 Product safety 291 recovery 237 Progenitor cells 103 Promotors prokaryotic 47 eukaryotic 47 Proteases Proteasome Proteome fingerprint Protooncogene Purine nucleosyl phosphorylase 185 68 216 65 This page has been reformatted by Knovel to provide easier navigation 355 Index terms Links Q Quality control 160 169 324 assurance 160 109 Qualification of equipment 229 R RAC 275 276 294 RCA 209 232 Radiotherapy 81 Raw materials 323 Receptors Regulatory aspects 281 framework 291 guidance 223 Retinoblastoma Retroviruses 336 Replication 185 Reproducability 147 315 Risk assessment 331 RNase 157 163 S Safety profile 277 Scientific reviews 293 Selective precipitation 101 103 Sepharose 104 This page has been reformatted by Knovel to provide easier navigation 356 Index terms Sequencing SOD Links 208 25 Sodium hydroxide 316 Sodium dodecyl sulphate 316 Sinapinic acid 206 Solid phase extraction 101 103 Somatic cells 245 gene therapy 299 tissue SOP 282 Spermine 103 Spermidine 103 Staph-epidermidus 183 Stoichimetric Strontium phosphate Suicide gene Sulphate glycoaminoglycans 65 140 22 115 245 Symptomatic therapies 245 Synthetic chromosomes 39 T Testing mycoplasma 191 potency 193 Thalassemia Thermophile 107 Thermopolymerase 109 This page has been reformatted by Knovel to provide easier navigation 357 Index terms Links Thyamidine kinase 65 synthase 65 Thymidine nucleoside 20 Thyroidine kinase 20 TIL 18 TNF 18 Transcription 45 65 Transduction 254 Transferrin 248 Transfiction 135 Transgene Transglutamide 265 Translation 45 Translocation 52 Transmission electron microscopy Tumour necrosis factor TSE 60 210 179 187 74 197 U Ultracentrifugation 234 Ultrafiltration 234 Unit operation 229 V Vaccination curative 155 genetic 155 This page has been reformatted by Knovel to provide easier navigation 358 Index terms Links Vaccination (Continued) veterinary DNA 157 Validation cleaning 229 234 237 viral clearance 234 gene therapy 223 process 228 Vector AAV 255 adenoviral 85 195 adenovirus 231 297 adeno-associated 298 chimeric 258 cosmid 54 envelope 142 intact 213 lentivirus 297 non-viral 137 260 polymeric 218 retrovirus 296 system 157 Virus adeno 253 adeno-associated 195 adventitious 192 alpha 268 artificial 256 bank preparation 232 This page has been reformatted by Knovel to provide easier navigation 359 Index terms Links Virus (Continued) master 232 safety 299 working 232 concentration 212 hermes simplex 256 onyx 255 mastadino 253 murine leukaemia 250 rhino virus 250 Sind bis 268 testing 172 Validation 313 323 Vogelstein 67 VP 22 261 This page has been reformatted by Knovel to provide easier navigation ... passed on to the next generation This type of gene therapy is known as somatic gene therapy and is in contrast to the concept of germ line gene therapy (which would involve a gene being introduced... Germ line gene therapy Introduction of genetic material into the egg cells or sperm cells of an individual such that the gene will also be passed on to the next generation Ex vivo gene therapy... of the patient's cells, introduction of therapeutic genetic material into these cells and reintroduction of these cells into the patient In vivo gene therapy Direct injection of therapeutic gene