CELL TECHNOLOGY FOR CELL PRODUCTS

Cell Technology for CellProductsProceedings of the 19th ESACT Meeting,Harrogate, UK, June 5-8, 2005

Edited by

Rodney SmithCTM BioTech Ltd, Babraham Cambridge, UK

A C.I.P. Catalogue record for this book is available from the Library of Congress.

ISBN-10 1-4020-5475-0 (HB)ISBN-13 978-1-4020-5475-4 (HB)ISBN-10 1-4020-5476-9 (e-book)ISBN-13 978-1-4020-5476-1 (e-book)

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Contents 19th ESACT Meeting Committees

Acknowledgements

CHAPTER I: PERSPECTIVES OF THE BIOTECH INDUSTRY

Tony Bradshaw CHAPTER II: TRANSCRIPTION TO SECRETION A Novel AIR-induced Gene Expression System in HEK.EBNA Cells and its Applications 11–18 Towards Stronger Gene Expression - a Promoter's Tale 19–28

Improving the Expression of a Soluble Receptor: Fc fusion Protein in CHO Cells by Coexpression with the Receptor Ligand 29–39

Growth, Apoptosis and Functional Genomics Analysis of

Microarray Analysis of Metabolically Engineered NS0 Cell Lines Producing Chimeric Antibody 47–52

List of Participants xxxiii

xxiii ESACT Executive Committee xxv Sponsors xxvii Companies in Trade Exhibition xxix

xxxi

v

N.S. Werner, W. Weber, M. Fussenegger, and S. Geisse

G.W. Lee, J.K. Fecko, A. Yen, D. Donaldson,

S. Kalwy, J. Rance, A. Norman,

CHO-K1 Over-Expressing Telomerase 41–45

and M. Leonard

and R. Gay

G. Khoo, F. Falciani, and M. Al-Rubeai

and M. Al-Rubeai F. Crea, D. Sarti, F. Falciani,

C. Wood, S. Tobler, S. Vunuum, Y. Luo,

The UK bioprocessing sector 3–7

vi

Optimizing Medium for Transient Transfection 53–55

Establishment of Recombinant CHO Cell Lines Under Serum-free Conditions 57–61

63–65

Characterisation of the Induction of Cytochrome p450 Enzymes in Primary Cultures of Human Hepatocytes 67–70

71–85

87–89 T. Hamasaki, S. Aramaki, T. Imada, K. Teruya, S. Kabayama,

Molecular Mechanism of TAK1-Induced Repression of hTERT Transcription 91–93

Information on High Producing Recombinant CHO Cell Lines 95–98

Redox Regulation by Reduced Waters as Active Hydrogen

S. Shirahata, Y. Li, T. Hamasaki, Z. Gadek, K. Teruya,

Novel Cell-based Assay for the Detection of Murine type I IfN

M. Maura, Y. Katakura, T. Miura, T. Fujiki,

type 2 Diabetes 99–101 Donors and Intracellular ROS Scavengers for Prevention of

Z.-H. Geng, W. Nudson, L. Davis, S. Luo, and K. Etchberger

M. Kuchenbecker, B. Rehberger, M. Schriek, J. Gackowski, and R. Fehrenbach

M. Bollati-Fogolín, and W. Müller

beta-Interferon in CHO Cells M. Spearman, J. Rodriguez, N. Huzel,

Effect of Culture Conditions on Glycosylation of Recombinant

K. Sunley, and M. Butler

Y. Katakura, K. Otubo, S. Morisawa, and S. Shirahata

H. Shiraishi, and S. Shirahata

C. Lattenmayer, M. Loeschel, K. Schriebl, E. Trummer,

Analysis of Genetic Parameters in Order to Get More

K. Vorauer-Uhl, D. Mueller, H. Katinger, and R. Kunert

S. Kabayama, K. Otsubo, S. Morisawa, Y. Ishii, and Y. Katakura

Reduced Water/platinum Nanoparticles in Cancer Cells

Contents

T.L. Freeman, A.P. Chadwick, M.S. Lennard,

Induction of Caspase-3-dependent Apoptosis by Electrolyzed

C.M. Martin, R.G. Turcan, and D. Blond

vii

Transcription Profiling of Different Recombinant CHO Clones Based on Cross-species Microarray Analysis 103–106

Adaptation without Cell Division? 107–110

Release of Plasmid-DNC from PEI/DNA Particles 111–115

Efficient Cloning Method for Variable Region Genes of Antigen Specific Human Monoclonal Antibody 117–119

Design Approach Regarding Humanization and

121–123

The Role of the Sumoylation Pathway in Transient and Stable Recombinant Protein Expression in Chinese Hamster Ovary Cells 129–132

Effect of Different Materials Used in Bioreactor Equipments on Cell Growth of Human Embryonic Kidney (HEK293) Cells Cultivated in a Protein-free Medium 133–136

Antibody Production by GS-CHO Cell Lines Over Extended Culture Periods 137–140

A.J. Porter, A.J. Dickson, L.M. Barnes, and A.J. Racher

E. Trummer, W. Ernst, D. Müller, K. Schriebl, C. Lattenmayer, K. Vorauer-Uhl, R. Kunert, H. Katinger, and F. Hesse

K.L. Astley and M. Al-Rubeai

M. Bertschinger, A. Schertenleib, G. Backliwal, M. Jordan, and F.M. Wurm

S. Matsumoto, Y. Katakura, M. Yamashita,

Functionality of an Anti-CD18 Monoclonal Antibody

Y. Aiba, K. Teruya, and S. Shirahata

C.A. Caldas, D.P. Souza, M.T.A. Rodrigues, A.Q. Maranhão,

for the Production of the Cancer Testis Antigen NY-ESO-1 125 128

A.M. Moro, and M.M. Brigido

D.L. Hacker, L. Cohen, N. Muller, H.-P. Thanh, E. Derow, G. Ritter,

Development of a Transient Mammalian Expression Process

and F.M. Wurm

M. Bertschinger, D. Hacker, and F. Wurm

P. Aizawa, G. Karlsson, C. Benemar, V. Segl, K. Martinelle, and E. Lindner

–

Contents

viii

Growth Promoting Effect of a Hydrophilic Fraction of the Protein Hydrolysate Primatone on Hybridoma Cells 141–146

Effect of the Scaffold in Maintenance of Suitable Chondrcyte Morphology in Cartilage Regeneration 147–151

Metabolic Changes Occuring During Adaptation to Serum Free Media 153–156 The Expression of the neo gene in GS-NS0 Cells Increases their Proliferative Capacity 157–162

CHAPTER III: THERAPEUTIC CELL ENGINEERING The Therapeutic Potential of Human Embryonic Stem Cells 165–184

Investigations of Murine Embryonic Stem Cell Maintenance by Analyses of Culture Variables and Gene Expression 185–189

New Electrofusion Devices for the Improved Generation of Dendritic Cell-tumour Cell Hybrids 207–216

J.J. Cairó, and F. Gòdia

S.H. Cedar, M.J. Patel, S.J. Pickering, P.R. Braude, and S.L. Minger

M. Schomberg, C. Hakemeyer, H. Büntemeyer,

J. Rodríguez, X. Rubiralta, J. Farré, P.J. Fabregues,

L. Stiens, and J. Lehmann

F. Veraitch and M. Al-Rubeai

S. Denby, F. Baganz, and D. Mousdale

J.M. Piret, C.H. Glover, M. Marin,

Generation of Cardiomyocytes Derived from Mouse

M.A.S. Chaudhry, C.J. Eaves, B.D. Bowen,

ESC in a Scale-up System 191–195

Expansion of Undifferentiated Human Embryonic Stem Cells 197–205

S. Niebruegge, H. Ebelt, M. Jungblut, H. Baer, M. Schroeder,

R.K. Humphries, C.D. Helgason, and J. Bryan

R. Zweigerdt, T. Braun, and J. Lehmann

A.B.H. Choo, J. Crook, and S.K.W. Oh

J. Schaper, H.R. Bohnenkamp, and T. Noll

J.M. Aguilera, J. Barrachina, F. Granell, J. Nebot,

Contents

ix

Influence of c-Myc and bcl-2 CO-expression on Proliferation, Apoptosis and Survival in CHO Cultures 217–220

Ex Vivo Expansion of Hematopoietic Stem Cells Using Defined Culture Media 221–224

Development of A Novel Serum-free Cryopreservative Solution 225–231

Hepatic Cell Lines CultureD on Different Scaffolds and in Different Stages for Bioartificial Liver Systems 233–238

Comparison of Flask and Bag Systems in Cultivation of Dendritic Cells for Clinical Application 239–242 L. Macke, B. Wrobel, W. Meyring, H.S.P. Garritsen,

T-Lymphocytes Transduced with SFCMM-3 Vector in a Closed System 243–250

Influence of Down-regulation of Caspase-3 and

Producing Thrombopoietin 251–258

CHAPTER IV: GENE MEDICINE

Protein Secretion From Mammalian Cells 261–268

V. Ifandi and M. Al-Rubeai

J. Hartshorn, A.S.-H. Chan, Z.-H. Geng, L. Davis, S. Luo, J. Ni, and K. Etchberger

T. Toyosawa, M. Sasaki, Y. Kato, H. Yamada, and S. Terada

K. Kaito, Y. Narita, and S. Terada

B. Wörmann, M. Rhode, W. Lindenmaier, and K.E.J. Dittmar

C. Benati, R.S. Birolo,

Apoptotic Cell Death of Chinese Hamster Ovary Cells

S.L.S. Catamancio, M. Radrizzani,

CASPASE-7 by Sirnas on Sodium Butyrate-induced

C. Sendresen, and S. Toma

Y.H. Sung, S.H. Park, S.H. Kim, and G.M. Lee

B. Stern, C. Gjerdrum, S. Knappskog, L. Minsaas, S. Nylund, L.C. Olsen, L. Olsen,

UTRtech™: Exploiting mRNA Targeting To Increase

A. Tveit, E. Vollsund, J.E. Hesketh, E. Oveland, H. Ravneberg, C. Trösse,

A. Tauler, and I.F. Pryme

Contents

x

Semi-Synthetic Mammalian Gene Regulatory Networks 269–272

Acoustic Cell Processing for Viral Transduction or Bioreactor Cell Retention 273–278

Comparison of Different Transient Retroviral Transfection Systems 279–282

CHAPTER V: CELLS TO TISSUES Artificial Human Lymphnode 285–290

Single Cell Study in a Hydrogel 291–295

Predicting Tissue Assembly of Prostate Cancer Spheroids 297–302

Cultivating Cells of Different Origin for 3d Bone Constructs

K. Suck, C. Kasper, C. Hildebrandt, S. Diederichs, M. Fischer,

Guidance on Good Cell Culture Practice 313–315

In Vitro Vascularization of Human Microtissues 317–320

V.M. Gorenflo, P. Beauchesne, V. Tayi, O. Lara, H. Drouin,

B.P. Kramer, M. Fischer, and M. Fussenegger

J.B. Ritter, V. Chow, C. Sherwood, B.D. Bowen, and J.M. Piret

and B. Opalka V. Kleff, I. Rattmann, C. Ludwig, M. Flasshove, T. Moritz,

C. Giese and U. Marx

K. O’Connor, C. Vidulescu, S. Clejan,

Considering Physiological Conditions 303–311

H. Song, and M. Venczel

T. Scheper, and M. van Griensven

S. Coecke, M. Balls, G. Bowe, J. Davis, G. Gstraunthaler, T. Hartung, R. Hay, A. Price, O.-W. Merten, W. Stokes, L. Schechtman, and G. Stacey

J.M. Kelm, C.D. Sanchez-Bustamante, and M. Fussenegger

Contents

T. Braschler, R. Johann, U. Seger, H. Van Lintel, and P. Renaud

xi

Preparation of Salmon Atelocollagen Fibrillar Gel and Its Application to the Cell Culture 321–323

Propagation of USSC for Therapeutic Approaches 325–329

Bioreactor Based Production of Functional Conditioned Medium for the Propagation of Undifferentiated Human Embryonic Stem Cells 331–334

Large-scale Expansion of Cells for Use as Therapeutic Agents 335–338

Hematopoietic Stem Cell Expansion Medium Generates a Large Population of Functional Early and Late Progenitor Cells 339–341

In Vitro Bone Formation of Multipotential Human Mesenchymal Cells in Fixed Bed Perfusion Bioreactors 343–346

Influence of Basal Media Formulations and Calcium on

Cells In Vitro 347–351

BHRF-1 as a Tool for Genetic Inhibition of Apoptotis in Hybridoma Cell Cultures 355–361

N. Nagai, S. Yunoki, T. Suzuki, Y. Satoh, K. Tajima, and M. Munekata

J. Lamlé, H. Büntemeyer, A. Reinel, T. Menne,

A. Bauwens, H. Büntemeyer, S. Terstegge, T. Nottorf,

C. van den Bos, and J. Lehmann

and M. Al-Rubeai

S.L. Leugers, D.W. Allison, Y. Liang, C. Swiderski, G. Van Zant, and L.M. Donahue

N. Zghoul, M. van Griensven, J. Zeichen,

Growth and Directed Differentiation of Human Osteogenic

I. Majore, M.M. Hosseini, H. Mayer,

CHAPTER VI: PROTEIN PRODUCTS

and V. Jäger

K.E.J. Dittmar, M. Rohde, and V. Jäger

S. Juanola, J. Vives, J. G lvez, P. Ferrer, R. Tello, J.J. Cairo, and F. G dia

O. Brüstle, and J. Lehmann

EX VIVO Expansion of CD34+ Cells in Stemline. II

J. Melero-Martin, S. Santhalingam,

TM

áò

Contents

xii

Karyotype of CHO DG44 cells 363–366 D. Martinet, M. Derouazi, N. Besuchet, M. Wicht,

Microcarrier Cultures as Sub-process 367–369

Effect of Peptones and Study of Feeding Strategies in a

Human Monoclonal Antibody 371–374

Retinoic Acid Induced Up-regulation of Monoclonal Antibody Production 375–379

Media and Cell Manipulation Approaches to the Reduction

The Production of Human Growth Hormone 385–391

Preparation of Membrane Receptor Proteins Utilising

Factory" to Finalise the Product 393–396

Silk Protein Sericin Improves Mammalian Cell Culture 397–401

Design of Culture Media for Drosophila Melanogaster

J. Beckmann, and F.M. Wurm

S. Nagel-Heyer, S. Lünse, C. Leist,

CHO Based Fed-batch Process for the Production of a

R. Böer, C. Goepfert, and R. Pörtner

V. Chotteau, C. Wåhlgren, and H. Pettersson

of Adaptation Time in GS NS0 Cell Line 381–383

and Y. Sugimoto

P. Clee and M. Al-Rubeai

F. Anton, A. Tappe, C. Kasper,

Large Scale Micro-carrier Culture and a "Pearl Freezing

A. Loa, B.-U. Wilhelm, and T. Scheper

I. Jansson, T. Fröberg, A. Danielsson, and E. Lullau

S2 Cells Producing Recombinant G Glycoprotein

M. Sasaki, and H. Yamada

from Rabies Virus 403–413

S. Terada, N. Takada, K. Itoh, T. Saitoh,

A.L.L. Galesi, F.R.X. Batista, R.Z. Mendonça, C.A. Pereira, and A.M. Moraes

From Biopsy to Cartilage-carrier-constructs by Using

Y. Inoue, H. Kawahara, S. Shirahata,

Contents

xiii

Comparison of the Cultivation of Wild and Transfected Drosophila Melanogaster S2 Cells in Different Media 415–423

Low Temperature Culture Effects on sCR1 Productivity by rCHO Cells 425–429

Proteomic Characterization of Recombinant NS0 Cell

431–434

IFN-Nucleotide and Sugar Nucleotide Contents of CHO Cells

N. Kochanowski, F. Blanchard, E. Guedon, A. Marc,

Influence of the Adaptation Procedure and the Serum-free Conditions on the N-glycosylation Pattern of EPO Produced by CHO Cells 439–442

Stability and Cytogenetic Characterization of Recombinant CHO Cell Lines Established by Microinjection and Calcium Phosphate Transfection 443–446 M. Derouazi, D. Martinet, N. Besuchet, R. Flaction,

and F.M. Wurm Efficient Cloning Method for Variable Region Genes of Antigen Specific Human Monoclonal Antibody 447–449

Design Approach Regarding Humanization and Functionality of an Anti-CD18 Monoclonal Antibody 451–454

K. Swiech, A.L.L. Galesi, A.M. Moraes, R.Z. Mendonça,

Clones Obtained by an Integrated Selection Strategy

H. Matsuoka, C. Shimizu, and T. Takeda

A. Castillo, K.R. De La Luz, S. Victores,

Cultivated in Batch Culture 435–438

R. Cacan, F. Chirat, and J.-L. Goergen

A. Marc, and J.-L. Goergen F. Le Floch, B. Tessier, J.-M. Guillaume, P. Cans,

M. Wicht, M. Bertschinger, D. Hacker, J. Beckmann,

S. Matsumoto, Y. Katakura, M. Yamashita, Y. Aiba, K. Teruya, and S. Shirahata

C.A. Caldas, D.P. Souza, M.T.A. Rodrigues,

L. Rojas, Y. Rabasa, S. Gaskell, and R. Perez

C.A. Pereira, and C.A.T. Suazo,

Glycosylation Macroheterogeneity and Intracellular

A.Q. Maranhão, A.M. Moro, and M.M. Brigido

Contents

xiv

Development of a Transient Mammalian Expression Process for the Production of the Cancer Testis Antigen NY-ESO-1 455–458

Effect of Different Materials used in Bioreactor Equipments on Cell Growth of Human Embryonic Kidney (HEK293) Cells Cultivated in a Protein-free Medium

Antibody Production by GS-CHO Cell Lines Over Extended

Growth Promoting Effect of a Hydrophilic Fraction of the Protein Hydrolysate Primatone on Hybridoma Cells

Effect of the Scaffold in Maintenance of Suitable Chondrcyte Morphology in Cartilage Regeneration J. Rodríguez, X. Rubiralta, J. Farré, P.J. Fabregues, J.M. Aguilera, J. Barrachina, F. Granell, J. Nebot, J.J. Cairó, and F. Gòdia Metabolic Changes Occuring During Adaptation to Serum

The Expression of the neo Gene in GS-NS0 Cells

1000 Non-instrumented Bioreactors in a Week

459–462

Culture Periods 463–466

467–472

473–477

Free Media 479–482

Increases Their Proliferative Capacity 483–488

489–495

D.L. Hacker, L. Cohen, N. Muller, H.-P. Thanh, E. Derow, G. Ritter, and F.M. Wurm

K. Martinelle, and E. Lindner

A.J. Porter, A.J. Dickson, L.M. Barnes, and A.J. Racher

M. Schomberg, C. Hakemeyer, H. Büntemeyer, L. Stiens, and J. Lehmann

S. Denby, F. Baganz, and D. Mousdale

F. Veraitch and M. Al-Rubeai

M. Stettler, M.D. Jesus, H. Ouertatani-Sakouhi, E.M. Engelhardt, N. Muller, S. Chenuet, M. Bertschinger, L. Baldi, D. Hacker, M. Jordan, and F.M. Wurm

P. Aizawa, G. Karlsson, C. Benemar, V. Segl,

Contents

xv

Optimisation of Time-space-yield for Hybridoma fed-batch

Influenza Vaccines – Challenges in Mammalian Cell

Towards pCO2 -optimised Fermentations – Reliable Sampling, pCO2 Control & Cellular Metabolism

Bioreactor and Stirred-Tank Reactor for Parvovirus

Serum-free Transient Expression of Antibodies

Monitoring Cell Physiology in Influenza Vaccine

NucleoCounter – An Efficient Technique for the Determination

On-line Biomass Monitoring in Cell Culture with Scanning

Cultures with an Adaptive Olfo-controller 497–501

509–518

Vaccine Production 519–522

Production by Flow Cytometry 531–533

of Cell Number and Viability in Animal Cell Culture Processes 535–538

Radio-frequency Impedance Spectroscopy 539–541

R. Pörtner, B. Frahm, P. Lane, A. Munack,

Culture Technology 503–508

K. Kühn, and V.C. Hass

Y. Genzel, J. Schulze-Horsel, L. Möhler, Y. Sidorenko, and U. Reichl

A. Jockwer, C. Klinger, J. Gätgens, D. Eisenkrätzer, and T. Noll

B. Hundt, S. Straube, N. Schlawin, H. Kaßner, and U. Reichl

K. Russ, P. Schlenke, H. Huchler, H. Hofer,

Using 40 Tray Cell Factories 527–530

M. Kuchenbecker, and R. Fehrenbach

J. Schulze-Horsel, Y. Genzel, and U. Reichl

Cultivation of E-FL in Perfusion Mode Using Wave®

P. Schlenke, K. Langnickel, T. Grob, F. Kohne, H. Hofer, U. Brändle, K. Russ, and R. Fehrenbach

D. Shah, P. Clee, S. Boisen, and

J.P. Carvell and C. Cannizzaro

M. Al-Rubeai

Repeated Batch Fermentation Using a Viafuge Centrifuge 523–526

Contents

xvi

The Use of a Mobile Robot for Complete Sample Management

A. Bock and U. Reichl Optimization of Culture Medium with the Use of

P. Hinterleitner, P. Ganglberger, B. Obermüller, B. Antes,

Serum-free Influenza Vaccine Production with MDCK Cells

Flow Cytometry as a Tool for Process Monitoring of

in a Cell Culture Pilot Plant 543–547

Protein Hydrolysates 553–557

559–562

563–565

567–569

Virus-cell Systems 571–573

575–577

579–581

M. Wojtczyk, R. Heidemann, K. Joeris,

Closed Loop Control of Perfusion Systems in High-density

C. Zhang, M. Burnett, A. Knoll,

Cell Culture 549–551

and K. Konstantinov

of Mab Ign 311 in Small Scale Experiments

P. Ganglberger, B. Obermüller, M. Kainer, P. Hinterleitner, O. Doblhoff, and K. Landauer

O. Doblhoff, and K. Landauer

Y. Genzel, M. Fischer, R.M. Olmer, B. Schäfer, C. Best, S. König, B. Hundt, and

Rotary Bioreactor for Recombinant Protein Production

U. Reichl

S. Navran

K.S. Sandhu and M. Al-Rubeai

B. Isailovic, R. Hicks, I.W. Taylor, A. Nienow, and C.J. Hewitt

J. Hartshorn, F. Guehenneux, K. Kolell, Z.-H. Geng, E. Atarod, T. Mohabbat, C. McCormick, P. Sutrave,

Understanding Metabolic Needs of EB14 Cells in Culture

C. Hernandez, M. Avery, S. Luo, M. Mehtali, and K. Etchberger

Optimization of a Fed-batch Strategy for Production

in Wave-bioreactor and 5L-stirred Tank Bioreactor

The Use of Multi-parameter Flow Cytometry for Characterisation of Insect Cell-baculovirus Cultures

Contents

xvii

Using Genomic Tools for the Identification of Important Signaling Pathways in Order to Facilitate Cell Culture

Plant Peptones: Nutritional Properties Sustaining

Monitoring of Extracellular TCA Cycle Intermediates

Comparison of Different Design Configurations of a 1-L Multi-bioreactor Station for Cell Culture

Ovary Cell Lines 595–597

599–601

in Mammalian Cell Culture 603–605

607–607

and Acceleratostat Experiments 609–609

Cell Culture Optimisation 611–611

Process Development 613–617

K. Kolell, J. Padilla-Zamudio, B. Schuchhardt, S. Gilliland, S. McNorton, B. Dalton, S. Luo, and K. Etchberger

D.K. Fong, S.L. Leugers, M.L. Hoepner,

N. Lin, J.L. Beckmann, J. Cresswell, J.S. Ross, B. Delong,

E. Olmos, N. Fischbach, and A. Marc

Z. Deeds, and M.V. Caple

J. Mols, N. Cavez, S. Agathos, and Y.-J. Schneider

J.B. Ritter, Y. Genzel, and U. Reichl

G. Wilson and A. Jørgensen

R.B. Nielsen and G. Wilson

G. Wilson

H. Schütte, U. Kolrep, L. Franck, and R. Wagner

and L.M. Donahue

Computational Fluid Dynamics Applied to Laboratory Scale

D.W. Allison, K.A. Aboytes, T.K. Johnson,

Cell Culture Reactors for the Prediction of Hydrodynamic Stress 587–590

A Comparison of the SixFors Fermenter System

Comparison of Cultivations of CHO Cell Line in Chemostat

and Lab Scale Fermenters For Clone Selection

Virus Production in Vero Cells Using a Serum-free Medium 583–585

Medium Development 591–593

Evaluation of a Novel Micro-bioreactor System for

Development and Application of An Animal-component

Recombinant Protein Secretion by CHO Cells

Free Single-cell Cloning Medium for Chinese Hamster

Contents

xviii

The Effect of Genetically Modified Insulin Growth Factor on Cell Proliferation and Adenovirus Productivity

for Manufacturing of Mammalian Cell Based

Purification of Rabies Virus Produced on Vero Cells Using

Human Growth Hormone (HGH) Purification From

Process Development for a Veterinary Vaccine Against

Working Towards a Chemically-defined Replacement

Effect of Vitamins or Lipids Addition on Adenovirus

Observation and Analysis of Lab Scaled Microcarrier Cultivation by In-situ Microscopy with Image

and T. Scheper

in Serum Free Medium 619–622

Biopharmaceuticals 623–628

Chromatography Techniques 629–634

Chromatographic Media 635–641

Heartwater Using Stirred Tanks 643–648

Processing Tools 657–661

A. Buckler and M. Al-Rubeai

S. Langhammer, M. Riedel, H. Bushnaq-Josting,

CHO-Cell Culture Supernatant Utilizing Macroporous

R. Brecht, R. Pörtner, and U. Marx

M. Chtioui, K. Trabelsi, and H. Kallel

J.-G. Walter, A. Tappe, C. Kasper, R. Zeidler, O.-W. Reif, and T. Scheper

I. Marcelino, M.F.Q. Sousa, A. Amaral, C. Peixoto, C. Verissimo, A. Cunha,

for Hydrolysates 649–651

M.J.T. Carrondo,

Z.W. Deeds, C.S. Updike, B.J. Cutak,

Production at High Cell Densities 653–656

and M.V. Caple

T.B. Ferreira, M.J.T. Carrondo, and P.M. Alves

G. Rudolph, A. Gierse, P. Lindner, C. Kasper, B. Hitzmann,

Evaluation of a Scalable Disposable Bioreactor System

and P. M. Alves

Contents

xix

Rapeseed Peptidic Fractions as Medium Additives in

B. Farges, B. Tessier, I. Chevalot, C. Harscoat, S. Chenu,

Influence of Rapeseed Protein Hydrolysis Conditions on Animal Cell Growth in Serum-free Media Supplemented

G. Chabanon, I. Chevalot, B. Farges, C. Harscoat, S. Chenu, J.-L. Goergen, A. Marc, and I. Marc Rapid Media and Process Optimization Using DOE:

Developing Chemically Defined Media Through DOE: Complete Optimization with Increased Protein Production

Bioreactor Process Strategies for rNS0 Cells

The Efficient Induction of Human Antibody Production in in Vitro Immunization by CPG Oilgodeoxynucleotides

Rapid Culture Media Optimization to Gain Multiple Fold Increase in Recombinant Protein Production in

Strategies for Large Scale Production

M. Pohlscheidt, U. Langer, B. Bödeker, D. Paulsen,

Proteomic Investigation of Recombinant CHO Cells

J.R. Cresswell, J.M. Zobrist, J. Wildsmith, N. Lin,

Animal Cell Bioreactors 663–665

with Hydrolysates 667–669

A Case Study 671–682

693–695

697–701

of Parapoxvirus Ovis NZ-2 707–710

in High Density Culture 711–713

I. Marc, J.L. Goergen, and A. Marc

S. Holdread, J.H. Kim, T. Oshunwusi,

in Less than 8 Months 683–691

J. Kuchibhatla, C. Hunt, and J.W. Brooks

K. Hammett, J. Kuchibhatla, C. Hunt, S. Holdread, and J.W. Brooks

D. Fernandez, J. Walowitz, W. Paul, L. Tescione, and S. Gorfien

Y. Aiba, M. Yamashita, Y. Katakura,

Two CHO Cell Lines 703–705

S.-E. Matsumoto, K. Teruya, and S. Shirahata

S.D. Storms, A. Chen, J. Yu, and T. Fletcher

H. Rübsamen-Waigmann, U. Reichl, H.J. Henzler, and H. Apeler

R. Valdes-Camin, and M. Caple

Contents

xx

Comparative Proteomic Study of Mechanisms Involved in the Expression of Recombinant Monoclonal Antibody

Monitoring Nutrient Limitations by Online Capacitance Measurements in Batch & Fed-batch CHO Fermentations

Characterisation of Shake Flasks for Cultivation of

Manufacturing of Lots of Human T-Lymphocytes

The Effects of Cell Separation with Hydrocyclones

Cultivation of Vero Cells on Microporous and Macroporous

715–721

723–726

Animal Cell Cultures 727–731

for Cell Therapy 733–737

739–744

Membrane Adsorbers 749–752

L.E. Rojas, K.R. de la Luz Hernández, S. Victores, L. Castellanos, S. Gaskell, A. Castillo, and R. Pérez

and G. Schmid S. Ansorge, G. Esteban, C. Ghommidh,

G. Jänicke, C. Sauter, R. Bux, and J. Haas

N.D. Bernardi, S. Trasciatti, A. Orlandi, F. Gaspari, I. Giuntini, T. Bacchi, S. Castiglioni,

Oxygen Control in Static Cell Cultures

E. Muru, M. Galagano, C.D. Mattei, L. Cavenaghi, and M.L. Nolli

N.D. Bernardi, E. Schwander, A. Orlandi, I. Tano, S. Castiglioni, E. Muru, F. Gaspari, M. Galgano, C.D. Mattei, L. Cavenaghi,

on the Viability of CHO Cells 745–748

and M.L. Nolli

R.C.V. Pinto, R.A. Medronho,

Erythropoietin Purification Employing Affinity

and L.R. Castilho

M.C.M. Mellado, D. Curbelo, R. Nobrega,

Microcarriers 753–755

and L.R. Castilho

M.C.O. Souza, M.S. Freire, and L.R. Castilho

in Non Amplified NS0 Myeloma Cell Line

Contents

xxi

CFD-Aided Design of Hollow Fibre Modules for Integrated Mammalian Cell Retention and Product

Comparison of Manufacture Technologies for RAV12 Monoclonal Antibody in Production Medium

A New Host Cell-line for the Fast, Reproducible and Efficient Production of Biopharmaceuticals

Analysis and Modeling of the Metabolism of Lepidopteran

Growth of Mammalian and Lepidopteran Cells on BioNOC® II Disks, a Novel Macroporous Microcarrier

Production of Human Growth Hormone in a Mammalian Cell High Density Perfusion Process T. Nottorf, W. Hoera, H. Buentemeyer, S. Siwiora-Brenke, A. Loa, and J. Lehmann

Proteomic Characterization of Recombinant NS0 Cell

Purification 757–760

761–763

769–771

Clones Obtained by an Integrated Selection Strategy 773–776

Cell Lines 777–780

781–784

789–793

R.L. Machado, A. Figueredo, D.G.P. Carneiro, L.R. Castilho, and R.A. Medronho

Biopharmaceuticals in CHO-Cells 765–767 S. Zahn, K. Abst, N. Palmen, S. Schindler, and A. Herrmann

B. Greulich, H.F. Abts, S. Zahn, and A. Herrmann

L. Rojas, Y. Rabasa, S. Gaskell, and R. Perez

J.-C. Drugmand, Y.-J. Schneider, and S. Agathos

J.-C. Drugmand, J.-F. Michiels, S. Agathos, and Y.-J. Schneider

J. Keenan, D. Pearson, and M. Clynes

Containing No Animal Derived Proteins

A. Castillo, K.R. De La Luz, S. Victores,

Screening Strategies for Iron Chelators in Serum Free Media 785–788

I. Shackel, A. Bass, A. Brewer, P. Brown,

Highly Efficient Serum-free Production of

M.C. Tsao, and L.W.S. Chang

Contents

xxii

Development of an Integrated Qualitative and Quantitative Approach to Study Expression-Secretion Bottlenecks of Mammalian cDNAs in

Rotating Bed Bioreactors: a Novel Way to Natural Proteins by Tissue-like Cultivation of Mammalian Cells

Minireactors for Animal Cell Culture: Monitoring of

Air-lift Bioreactor Improvement of Scale-up Linearity Between Shake Flasks and 25-L Scale in the Insect

HEK293 EBNA Cells 795–798

799–801

803–806

Cells–baculovirus Expression System 807–809

L. Glez, C.A. Power, T. Battle, and G. Feger

Cell Concentration and Cellular Activity

H.P.A. Hoffmeister

F. Bollin, C. Losberger, L. Chevalet,

R. Bragós, J.J. Cairó, and F. Gòdia

and T. Battle

A. Soley, J. Gálvez, E. Sarró, M. Lecina, A. Fontova,

Contents

Name Index

Subject Index

811 816 –

– 81 7 821

19th ESACT Meeting Committees

Organising Committee

Rodney Smith (Chair) CTM BioTech Ltd., UK Fiona Godsman (Trade Fair/Sponsorship) BioReliance, UK

Magellan Instruments Ltd., UK Tracey Sambrook (Social Arrangements) CTM BioTech Ltd., UK Glenda Bland (Conference Manager) Global Meeting Planning

Scientific Committee

Dr. Rod Smith (Chair) CTM BioTech Ltd., UK Prof. Mohammed Al-Rubeai University of Dublin, Ireland Prof. Dr. Martin Fussenegger ETH Zürich - Institute of Biotechnology,

SwitzerlandProf. Dr. Ing. Jürgen Lehmann Universität Bielefeld, Germany Prof. James M. Piret University of British Columbia, Canada Prof. Glyn Stacey National Institute for Biological

Standards and Control, UK Dr. David Venables BioReliance, UK

Poster Award Committee

Nathalie Chatzissavidou Biovitrum, Sweden Paula Marques Alves ITQB/IBET, Portugal

xxiii

John Bonham-Carter (Trade Fair/Sponsorship)

ESACT Executive Committee

Otto-Wilhelm Merten, Chairman Généthon III, France

Alain Bernard, Secretary Serono Biotech Center, Switzerland

Martin Fussenegger, Treasurer ETH- Hönggerberg , Zurich, Switzerland

Rodney Smith, Meeting Chair CTM BioTech, UK

Francesc Gòdia Universitat Autonoma de Barcelona, Spain

Hansjörg Hauser GBF, Germany

Stefanos Grammatikos Boehringer Ingelheim Pharma, Germany

Florian Wurm EPFL, Switzerland

Christophe Losberger, Web Master Serono Pharmaceutical, Research Institute, Switzerland

Steve Oh, Newsletter Chief Editor Bioprocessing Technology Institute, Singapore

Merlin Goldman, Newsletter Co-Editor Wyeth Biopharma, Ireland

Bryan Griffiths ESACT Secretariat, UK

xxv

Sponsors

ESACT and the Organising Committee wish to thank the following companies for their generous support.

Aber Instruments

AstraZeneca

Aventis Pasteur SA

ChemoMetic A/S

Covance

CTM BioTech Ltd.

DSM Biologics

GE Healthcare

Genetech

Genzyme Flanders NV

Invitrogen

JRH Europe

Magellan Instruments

Merck & Co., Inc., USA

Merck Research Laboratories

Rütten Engineering AG/Ltd.

Sartorius BBI Systems GmbH

Serono

Sigma Aldrich

Wave BioTech

xxvii

Companies in Trade Exhibition

Aber Instruments Ltd.

Applikon. BV/AppliSens

Areta International s.r.l.

Bayer HealthCare AG

Bayer Technology Services GmbH

BDD Diagnostic Systems Europe

Beckman Coulter

Bioengineering AG

BUA/BioProcess UK

BioProcess Engineering Services Ltd.

BioProcess International

BioProcessors Corporation

Bioquell

BioXplore

Blue Membranes GmbH

Broadley Technologies Ltd

Cambrex Bio Science

Celliance

Cellon SA

Celonic GmbH

Genetix Ltd.

Greiner Bio-One Ltd.

Guava Technologies

Helena Biosciences

Henogen

HyClone/Perbio Science

Infors UK Ltd./Hi Tec-Zang

Innovatis AG

Invitrogen

Irvine Scientific

J M Separations BV

JRH Europe

Kerry Bioscience

Lonza Biologics plc

Magellan Instruments

Moregate TCS Ltd

New Brunswick Scientific (UK) Ltd

NewLab BioQuality AG

NUNC A/S

PAA Laboratories GmbH

xxix

Companies in Trade Exhibition

Charter Medical Ltd.

Chemicon Europe

ChemoMetec A/S

Cobra BioManufacturing

Consolidated Polymer Technologies (C-Flex)

Corning Life Sciences

Covance

CTM BioTech Ltd.

DASGIP AG

Delta Biotechnology Ltd.

Eden Biodesign

European Collection of Animal Cell Cultures (ECACC)

Evotec Technologies

GE Healthcare

Genetic Engineering News

Pall Life Sciences

Panacea

Polyplus Transfection

Quest Biomedical

Rütten Engineering

Sartorius BBI Systems GmbH

Sigma Aldridch

SLS

Stedim S.A.

Tecan Deutschland GmbH

The Automation Partnership

The Williamsburg BioProcessing

Wave Biotech AG

Wave Europe Pvt. Ltd.

Xenova Biomanufacturing

YSI (UK) Limited

xxx

Acknowledgements

In the planning, organising and running the meeting a great number of people have been involved. The outcome was a successful meeting and these proceedings of the science presented. The 19th ESACT Meeting built on the success of the previous ones and lays the foundation for the 20th and beyond.

In making the meeting so successful I would like to acknowledge and thanks the following people for their effort hard work and most of all friendship over the period. These people provided immense support to me during the events and I shall never forget that.

My personal thanks must go to Fiona Godsman for staying on with the Trade Organisation and whose invaluable comments were appreciated and hopefully listened too. John Bonham-Carter (Trade Organisation) who was so keen in Tylosand to be involved and relished the chance of developing his (sometimes wild) ideas on the Harrogate meeting. Glenda Bland whose efforts in keeping us on track and focussed was extremely well received. Tracey Sambrook for her advice support and constant council on how the meeting was destined to be a success.

These people had the vision and energy to ensure that no part of the Harrogate meeting was forgotten or left out. I owe them the greatest of thanks for being part of the team.

I would also like to thanks the other people who helped in laying down the science for the community to ensure the programme was balanced and thought provoking. The main scientific committee comprising James Piret, Glynn Stacey, Martin Fussenegger, Mohammed Al-Rubeai and Jürgen Lehmann. We should not forget the organisers of the now ever present Poster award and my thanks go to Nathalie Chatissavidou and Paula Alves for the excellent organisation and selection of some truly exciting posters.

xxxi

The biggest acknowledgment and thanks must go to the delegates and all those scientists that wanted to participate in some way to the meeting. Without your contributions the meeting would not be the same.

A very big Thank you to you all.

Rodney Smith

xxxii Acknowledgements

List of Participants

Prof. Adriano Aguzzi, Institute of Neuropathology, University Hospital of Zurich, Schmelzbergstrasse 12, Zurich CH-8091, SWITZERLAND, Telephone: +4112552107, Email: adriano@pathol.unizh.ch

Mr. Yoshihiro Aiba, Laboratory of Cellular Regulation Technology, Department of Genetic Resources Technology, Kyushu University, Fukuoka, JAPAN, Telephone: +84 92 642 3046, Email: aiba@grt.kyushu-u.ac.jp

Mr. Peter Aizawa, Octapharma AB, Nordenflychtvägen 55 Stockholm 12137, SWEDEN, Telephone: +46 8 56643436, Email: peter.aizawa@octapharma.se

Mr. Franz Alig, Sartorius BBI Systems GmbH, Schwarzenberger Weg 73 - 79, Melsungen D-34212, GERMANY

Mr. Tony Allman, Infors UK Ltd., The Courtyard Business Centre, Dovers Farm, Lonesome Lane, Reigate Surrey RH2 7QT, UK

Prof. Mohammed Al-Rubeai, Department of Chemical & Biochemical Engineering, Falculty of Engineering & Architecture, University College Dublin, Belfield, Dublin 4, Ireland, Email: m.al-rubeai@ucd.ie

Dr. Paula Alves, ITQB/IBET, Apartado 12, OEIRAS 2781-901, PORTUGAL, Telephone: + 351 21 4469421, Email: marques@itqb.unl.pt

Dr. Madawi Alwazzan, Flowgen Bioscience, Wilford Industrial Estate, Ruddington Lane, Nottingham NG11 7EP, UK, Telephone: +44 115 977 5494, Email: malwazzan@flowgen.co.uk

Dr. Hagit Amitai, Compugen Ltd., 72 Pinchas Rosen Street, Tel-Aviv 69512, ISRAEL, Telephone: +972 3 7658513, Email: hagita@compugen.co.il

Dr. Anette Amstrup Pedersen, Novo Nordisk A/S, Novo Alle, Bagsvared 2880, DENMARK, Telephone: +45 44 42 00 08, Email: aamp@novonordisk.com

Dr. Dana Andersen, Director, Late Stage Cell Culture, Genentech, Inc., 1 DNA Way - MS 32, South San Francisco, CA 94080, USA, Telephone: +1 650 225 4443, Email: andersen@gene.com

xxxiii

List of Participants

Mr. Jorn Sondrup Andersen, Rocket Management, Ganloseparken 16, Stenlose DK-3660, DENMARK, Telephone: +45 22 16 67 26, Email: sondrup@rocketmail.com

Mr. William Anderson, Synthecon, Inc., 8042 El Rio, Houston, TX 77054, USA, Telephone: +1 713 741 2582, Email: abill@synthecon.com

Dr. Carlo Andretta, Biospectra AG, Zuercherstrasse 137, Schlieren 8952, SWITZERLAND, Telephone: +41 44 7556700, Email: andretta@biospectra.ch

Mrs. Emilie Ansel, GlaxoSmithKline Biologicals S. A., rue de Institut 89, Bixensart 1330, BELGIUM, Telephone: +32 2 656 9890, Email: emilie.ansel@gskbio.com

Ms. Fabienne Anton, Insitut für Technische Chemie - Universitat Hannover, Callinstrasse 3, Hannover 30167, GERMANY, Email: anton@iftc.uni-hannover.de

Mr. Didier Argentin, Henogen, Rue des Professeurs Jeener et Brachet 12, Charleroi B-6041, BELGIUM

Mr. Paul Armstrong, Celliance, 5655 Spalding Drive, Norcross, GA 3009, USA

Mr. William Arts, New Brunswick Scientific B.V., Kerkenbos 1101, Nijmegen 6546 BC, THE NETHERLANDS, Telephone: +31 24 371 7600, Email: w.arts@nbsbv.nl

Dr. Soheil Asgari, Blue Membranes GmbH, Rheingaustrasse 190-196, Wiesbaden D-65203, GERMANY

Ms. Kelly Astley, 41 Wealdstone Drive, Lower Gornal, Dudley, West Midlands DY3 2QR, UK, Telephone: +441384 821 487, Email: kla266@bham.ac.uk

Mr. Alex Baker, Maverick DCM, Suite 12-16 Macquarie Chambers, 69-79 Macquarie Street, PO Box 2997, Dubbo 2831, AUSTRALIA, Telephone: +61 2 6885 1200, Email: ahanslow@maverickdcm.com

Mr. Osman Bal, Git Verlag, Roessler Str. 90, Darmstadt 64283, GERMANY

Mr. John Baldwin, Rütten Engineering Ltd., Industriestrasse 9, Stäfa 8712, SWITZERLAND, Telephone: +41 1 928 29 30, Email: info@rutten.com

Mr. Jean-Philippe Barbe, Sanofi Pasteur, 1541 av. M. Mérieux, Marcy l'Etoile 69280, FRANCE, Telephone: +33 4 3737 3294, Email: jean-philippe.barbe@sanofipasteur.com

Dr. Jacquiline Barnes-Locke, Helena Biosciences, UK

Dr. Barney Barnett, Biogen Idec, 5200 Research Place, San Diego 92122, USA, Telephone: +1 858 401 8650, Email: barney.barnett@biogenidec.com

Mr. Timothy Barrett, University College London, UK, Email: timothy.barrett@ucl.ac.uk

Ms. Natalie Baseley, European Collection of Cell cultures (ECACC), Health Protection Agency, Porton Down, Salisbury SP4 0JG, UK

Miss. Vera Basto, ECBio-R&D in Biotechnology, SA, Ap 98, Lab 4.11, Ed ITQB, Oeiras 2781-901, PORTUGAL, Telephone: +351214469466, Email: vera.basto@ecbio.com

xxxiv