ANIMAL CELL TECHNOLOGY Products of Today, Prospects for Tomorrow Editors R. E. Spier Department of Microbiology, University of Surrey Guildford, Surrey, UK J. B. Griffiths PHLS CAMR, Porten, Salisbury, Wilts, UK W. Berthold Dr Karl Thomae GmbH, Biberach an der Riss, Germany EUROPEAN SOCIETY p i U T T E R W O R TH FOR ANIMAL CELL TECHNOLOGY ■|E I N E M A N N THE 12th MEETING Butterworth-Heinemann Ltd Linacre House, Jordan Hill, Oxford 0X2 8DP Ό^Α member of the Reed Elsevier group OXFORD LONDON BOSTON MUNICH NEW DELHI SINGAPORE SYDNEY TOKYO TORONTO WELLINGTON First published 1994 © Butterworth-Heinemann Ltd 1994 All rights reserved. No part of this publication may be reproducde in any material form (including photocopying or storing in any medium by electronci means and whether or not transientyl or incidentalyl to some other use of this publication) without the written permission of the copyright holder except in accordanec with the provisions of the Copyright, Designs and Patenst Act 1988 or under the terms of a licence issued by the Copyright Licensing Agency Ltd, 90 Tottenham Court Road, London, England W1P 9HE. Applications for the copyright holders' written permission to reproduce any part of this publication shoudl be addressde to the publishesr British Library Cataloguing in Publication Data A catalogue record for this book is availabel from the British Library ISBN 0 7506 1845 0 Library of Congress Cataloguing in Publication Data A catalogue record for this book is availabel from the Library of Congress Printed in Great Britain by Redwood Books, Trowbridge, Wiltshire European Society for Animal Cell Technology ESACT is a socieyt of scientist,s engineesr and contributosr to progress in the area of animal cell culture and technology. ESACT has been founded to creaet a forum for the exchange of ideas on biological and engineering techniques to promoet knowledge and use of animal cells e.g. for the manufacturing of products. ESACT is a member of the EFB (European Federation of societies for Biotechnology) and assumse a role to further the understandin,g researhc and use of animal cells by organising regular international scientific meetings and communicatiosn to the membesr and to political bodies. ESACT MEETINGS ESACTs' sesquiennlia meetings are designed to give room for schedules presentation, sfor reporst and critical evaluation as well as informal discussio.n Time is also set aside for unschedulde dialogue among the limited number of attendese to allow for an 'in depth' discussion and brain storming on topics concerning animal cell technology. The proceedinsg of ESACT meetings are published. RECENT ACHIEVEMENTS of animal cell technology in: manufacturing of producst engineering of technical devices of cell handling engineering of instrumentatino for cell culture control and equipment engineering of cell systems as a source for producst and scientific researhc NEW APPLICATIONS of animal cells made possibel by genetci engineering or new technical concepst for: generating new producst e.g. novel or rare proteins generating in vitro test systems e.g. for toxicological assessmesn t generating new types of producst e.g. cells for gene therapy PROSPECTS of developmetn with impact on animal cell culture in: redefining medical applications extending the scope to product types other than vaccines or proteins opening the way for new approachse in researhc v ESACT Executive Committee (9/91 - 5/93) Chairman and Meeting Secretayr Wolf Berthold Thomae GmbH, Biberach, Germany Secretayr /Treasurer Caroline MacDonadl U Of Paisley, Paisley, UK Member David Broad Celltech, UK Member E. Coen Beuvery RIVM, Bilthoven, The Netherlands Member Bryan Griffiths PHLS CAMR, Salisbury, UK Member Hansjörg Hauser Institut Pasteu,r Paris, France Member Ray Spier U of Surrey, Guildford, UK Organising Committee Wolf Berthold Thomae GmbH, Biberach/Riss Ullrich Behrendt Boehringer Mannheim, Penzbegr Guy Berg Sartorius, Göttingen Mike Comer Boehringer Mannheim, Penzbegr Elisabeht Fraune B. Braun Diessel Biotech, Melsungen Hansjörg Hauser GBFmbH, Braunschwegi Jürgen Lehmann Universität, Bielefeld, Bielefeld Herbert G Miltenburger Technische Hochschule, Darmstadt Wolfgang Noe Thomae GmbH, Biberach/Riss William Werz Thomae GmbH, Biberach/Riss VI ESACT Executive Committee (9/91 - 5/93) Chairman and Meeting Secretayr Wolf Berthold Thomae GmbH, Biberach, Germany Secretayr /Treasurer Caroline MacDonadl U Of Paisley, Paisley, UK Member David Broad Celltech, UK Member E. Coen Beuvery RIVM, Bilthoven, The Netherlands Member Bryan Griffiths PHLS CAMR, Salisbury, UK Member Hansjörg Hauser Institut Pasteu,r Paris, France Member Ray Spier U of Surrey, Guildford, UK Organising Committee Wolf Berthold Thomae GmbH, Biberach/Riss Ullrich Behrendt Boehringer Mannheim, Penzbegr Guy Berg Sartorius, Göttingen Mike Comer Boehringer Mannheim, Penzbegr Elisabeht Fraune B. Braun Diessel Biotech, Melsungen Hansjörg Hauser GBFmbH, Braunschwegi Jürgen Lehmann Universität, Bielefeld, Bielefeld Herbert G Miltenburger Technische Hochschule, Darmstadt Wolfgang Noe Thomae GmbH, Biberach/Riss William Werz Thomae GmbH, Biberach/Riss VI ACKNOWLEDGEMENT The Organizing Committee of the 12th ESACT Meeting would like to acknowldge the following organisations and thank them for their generous support AMGEN GENENTECH Inc. CIBA-GEIGY Behringwerke AG Dr. Karl Thomae GmbH Sartorius AG Boeringer Mannheim GmbH and their interest in ESACT by helping with the conference material and participation in the trade show HYCLONE EUROPE LTD, JRH BIOSCIENCES PHARMACIA BIOPROCESS TECHNOLOGY QUALITY BIOTECH LTD, SIGMA CHEMIE GmbH and Abion GmbH Kluwer Amicon Life Technologies GmbH B. Braun Biotech Int. GmbH Macmillan Journasl Bibby Dunn Labortechnki GmbH Microbiological Associates Int. Ltd Biolntermediari Millipore CanSear International New Brunswick Scientific GmbH Cellon Sari Nune GmbH Celltech Biologies pic Oxford Glycosystems Centritech AB PAA Labor und Forschung GmbH Chemap AG Partee GmbH Costar GmbH Protein Performanec SA Dr. Bräutigam MTH Sarstedt ECACC Schäref System GmbH Elsevier Serva Feinbiochemiac KG Fisons Instrumenst SGi Gibco BRL Südmo Schleiche rAG GIT TCS Biologicals Ltd Greiner GmbH Tecnomaar Hazelton Europe UniSyn Technologies Intergen Co Europe Westfalia Separatro AG Institute Pasteur/TexCel l John Wiley Inveresk Researhc International xiii Presenec and future of ACT Presence and Future of Animal Cell Technology Chairman's introduction to the 12th ESACT Meeting by Wolf Berthold Dept Biotech process Dev. Dr.Karl Thomae GmbH, Biberach/Ris,s Germany Introduction It is the prerogative of the chairman of this meeting to spend a few minutes of your time to offer some thoughst and reasonings that have led the organization to "compose" this meeting in its present format. Focus on essential perspectives in medicinal products It may not be new, but sometimes very refreshing to take a step back to view the whole area within its frame of reference, before all the biotechnical details become overpowering. To gain this viewpoint we can try to look at Animal Cell Technology/Biotechnoloyg through the eyes of the non-Biotechnologis. tIt is useful to try to define the market place for the specific producst animal cells can offer. We could compaer biotechnology with any other industry in following a trail of questions that could just as well be applied to the world of the automobile. In that industry the customer can be identified as the car user and car buyer, with his intentions, needs and possibilities. Questions for a decision tree: Who needs the product ? What are the user's requirements ? What choice for competing products do they have or want ? Who actually pays for the product ? Who can (profitably) provide or manufacture the product ? Where is the best research for products ? ? No one today needs to be a mechanci to enjoy driving a car. But many car users have an opinion on cars, and their non-expetr opinions can have an effect on the car industry as a whole and detailsof cars down to the very basci elemenst of car mechanic.s These technical details, their feasibility and perfection have to be addressde by specialisst who may have little appreciation for the overall needs of the car user. However, their success and recognition may well depend on how well they are able to implemen tthe customesr "whims". The producst that Animal Cell Technology can produce has to be appreciatde by non- Biotechnologist.s Endusesr are found exclusiveyl in area of health care as medicianl W.Berthold 1993 3 Presenec and future of ACT products and diagnostic.s To date, no necessyit or desire on the one hand, nor any convincing innovation on the other hand has resulted in animal cells cultivated in vitro for any other , non-medicinal (commercia)l use. Cosmetics and food industry may be the closest to todays applications of animal cells. Medicinal producst do not originate from animal cells alone - far from it. Most medicinal products are not even from any biological production source. But there are some products, that can not be produced in any other way. The most important and most traditional applications are vaccines for animal and human pathogenci viruses. More recently, a number of proteins have become availabel for the benefti to patients. Future possibilities for ACT : The downside In the near future there will be some more and novel proteins (e.g. antibodies) that are under investigation now. On the other hand some protein producst may also disappear again. This included unfortunateyl also the antibody against endotoxin, that had been hailed in the previous ESACT meeting as a remarkabel achievemen. tIt may be somewhat consoling to animal cell technology, that the reason for withdrawal from the market had nothing to do with the manufacturing proces.s Todays mode of application of pharmaproteisn is nearyl alway by parenterla injection, because the proteins target for action lies beyond the immune barrier inside the body. Injectabel proteins produced outside the body may howeve,r be replaced when they will be generatde within the patien.t The medicinal product could be a coding gene in the form of a "naked" nucleic acid. The active ingredient could also be a a nucleic acid/"antisens"e or a chemical peptide mimetic interacting with specific gene regulatoyr proteins at the genome level of the patienst cells (suppressro gene regulation ?). This kind of drug would not require cells fermentation anymore. The active ingredient would be made by bacterai or chemical synthesi.s Future possibilities for ACT : The upside Another form of future medicin could also be life cells themsel.f In analogy to the recombinant production cell line that produces the protein of choice in a fermente.r These cells howeve,r could be applied directly to the patient as cells. This mode of somatci gene therapy or (tumor-)' vaccine requires animal cell technology. The number of cells required, their safeyt test profile and the handling of such cells emphasiez different aspecst of todays' recommendde procedures for minimizing biological risks as compared to production cells. Probably, much less cells are sufficient, their productivity may not need to be "high yielding". As special feature, they shoudl not irritate the immune system of the patien.t Hence, they must be human and show histocompatibilivt with the patien.t Very important: they shoudl not pose a risk of infection or unlimited growth. Therefore, the technical aspecst of this cell handling may differ considerabyl in scael and mode from todays large fermentation tank technology. The handling of human cells with the intention of "regrafting" has a long standing tradition for suspendde normal cells in blood banks. Extracorporal cell treatmen,t and cell purification is used for blood transfusion of red blood cells. More recentyl bone marrow (in cancer treatmen)t and fetal brain cells (in parkinson diseas)e has been regrafted.Thsi W.Berthold 1993 4 Presenec and future of ACT kind of proceduer lend itself to extracorporla modification of other cells like in T-cell gene therapy. Analogous to this developmen, tcells may be given back to a patient not as single-cell suspensino but as complex 3-dimensionla "tissue". First steps in that direction are (artificial) skin for burns, artificial liver for detoxification or perhaps pancreatci islet cells in diabete.s In all these cases the technical handling of cells outside the body destined to return into a patient poses the challenge of even a higher degree of prevention of fortuitous contamination or break of sterility of cells. These features are important new criteria for the technical developmetn and engineering of devices designed for special applications beyond todays bioreactor.s Shift of goals of technical engineering Outlining these developmenst I want to picture the changing character of animal cell technology. The technical achievemetn of modern large scael fermentation of animal cells has been the fruit of continued effort in the design of good fermentation vesselsThe progress was guided by a number of goals : - maintenanec of sterility for long periods of time (robustness, ) - gentle handling of cells, - adequaet and safe harvesting procedures and - hopefully high final yield of product and - economci improvemenst in process design. These have been the dominant targest of researhc in both the technical and the biological aspecst of cell fermention. The resulting fermentesr today may still be improved. However, further improvemenst in design may give rise to comparativeyl modest economci advantagse compared with yield improvemenst due to new and more efficient vector construction.s Even those improvemenst may become irrelevan,t if the whole treatment mode changes from the protein itself as product to nucleic acid or cells as produc.t Animal Cell technology should take these new areas into consideration. They are qualitatively related but different in detail and therefore, scientifically in both biology and engineering and possibly economically more rewarding than the refinement of very good bioreactors already available today. Impact of the non "biological" environment on ACT How can ESACT achieve its dedication to further the application of animal cells. I may be forgiven, to provocatively rank a number of non-biologica lrealities next to Animal- and Biotechnology. Nameyl such mondäne realities as the time and expensse needed to achieve the final marketabel product besides the researhc and product oriented activities in biological disciplines. 5 W.Berthold 1993 Presenec and future of ACT 1. Safeyt assessmen.t Asll necessayr non-biotech obligations have to be fulfilled.Evidence of safeyt not only for the patien,t has to be provided, but for the bystander during production and handling of the cell. This includes validation and GMP (Good Manufacturing Practice)s and for the monitoring of the environment in compliance with all genetci engineering laws. 2. A major hurdle can be the granting of an Authorization of use of a Production Facility for commercial pharmaceuticla producst ( Drug regulation )based on GMO's (Geneticalyl Modified Organisms() Enviromental regulation.) 3. The Application for Authorization of Marketing of a medicinal produc.t This can also involve governmentla agreemenst on prizing as well as on medicinal efficacy. 4.Legal and patent issues. All the legal aspecst of patent laws, that may be touched by the gene, the cell, the proces,s the equipment etc. has to settled to avoid involuntary infringement or litigation. Ironocally, it is the legal profession, that has gained some of the most substantila financial benefits from biotechnology up to date. These facets of reality in the maturation process of a biological finding to a marketabel product are located after animal cell technology and researhc problems have been solved. They have to be considered well in advance to be overcome and make a return on investment possibel at all.. THE HOPE AND PERSPECTIVE FOR A NEW WORTHWHILE PRODUCT IS THE MOTOR FOR THE MERRY-GO-ROUND OF MONEY CHANGING HANDS WITHIN THE WORLD OF BIOTECHNOLOGY ONLY. MOST IMPORTANT FOR A VIABLE AND PROSPEROUS ANIMAL CELL TECHNOLOGY IS THE ACHIEVEMENT OF SUCCESSFUL PRODUCTS THUS CREATING BENEFIT FOR PATIENTS AND CREDIBILITY FOR THE ECONOMIC ATTRACTION OF THIS RELATIVELY YOUNG TECHNOLOGY. Some of these hurdles have to be addressde in a political arena. An exampel of impact is the protracted public discussion on hypothetical risks of genetci engineering in Germany. The uncertainyt of regulations (time and costs) has invited industrial investment and researchesr to go elsewher.e Where (at least in their eyes) unnecessayr stumbling stones are not eating away from their already decreasign grant money or economci health. To promote biotechnology with recombinant cells requires quite some courage in many countries in Europe. And from todays situation, it will need even more enduranec and effort ro regani the level expertise and the momentum in todays international competition for progress in gene- and bio- technology. 6 W.Berthold 1993