Process scale bioseparations for the biopharmaceutical industry

573 Pages·2007·11.53 MB·English

by  Abhinav A. Shukla

DK3347_half-title 6/2/06 11:31 AM Page 2
PROCESS SCALE
BIOSEPARATIONS FOR THE
BIOPHARMACEUTICAL
INDUSTRY
edited by
Abhinav A. Shukla
Amgen Inc.
Seattle,Washington, U.S.A.
Mark R. Etzel
University of Wisconsin-Madison
Madison,Wisconsin, U.S.A.
Shishir Gadam
Merck & Co., Inc.
West Point, Pennsylvania, U.S.A.
© 2007 by Taylor & Francis Group, LLC
DK3347_Discl.fm Page 1 Thursday, February 2, 2006 11:12 AM
Published in 2007 by
CRC Press
Taylor & Francis Group
6000 Broken Sound Parkway NW, Suite 300
Boca Raton, FL 33487-2742
© 2007 by Taylor & Francis Group, LLC
CRC Press is an imprint of Taylor & Francis Group
No claim to original U.S. Government works
Printed in the United States of America on acid-free paper
10 9 8 7 6 5 4 3 2 1
International Standard Book Number-10: 1-57444-517-0 (Hardcover)
International Standard Book Number-13: 978-1-57444-517-6 (Hardcover)
Library of Congress Card Number 2006002480
This book contains information obtained from authentic and highly regarded sources. Reprinted material is
quoted with permission, and sources are indicated. A wide variety of references are listed. Reasonable efforts
have been made to publish reliable data and information, but the author and the publisher cannot assume
responsibility for the validity of all materials or for the consequences of their use.
No part of this book may be reprinted, reproduced, transmitted, or utilized in any form by any electronic,
mechanical, or other means, now known or hereafter invented, including photocopying, microfilming, and
recording, or in any information storage or retrieval system, without written permission from the publishers.
For permission to photocopy or use material electronically from this work, please access www.copyright.com
(http://www.copyright.com/) or contact the Copyright Clearance Center, Inc. (CCC) 222 Rosewood Drive,
Danvers, MA 01923, 978-750-8400. CCC is a not-for-profit organization that provides licenses and registration
for a variety of users. For organizations that have been granted a photocopy license by the CCC, a separate
system of payment has been arranged.
Trademark Notice: Product or corporate names may be trademarks or registered trademarks, and are used only
for identification and explanation without intent to infringe.
Library of Congress Cataloging-in-Publication Data
Process scale bioseparations for the biopharmaceutical industry /
Abhinav Shukla, Mark Etzel, and Shishir Gadam, editors.
p. ; cm. -- (Biotechnology and bioprocessing series ; 31)
Includes bibliographical references and index.
ISBN-13: 978-1-57444-517-6 (alk. paper)
ISBN-10: 1-57444-517-0 (alk. paper)
1. Proteins--Purification. 2. Cell culture. 3. Pharmaceutical biotechnology. 4. Chromatographic
analysis. 5. Fermentation.
[DNLM: 1. Proteins--isolation & purification. 2. Cell Culture Techniques. 3.
Chromatography--methods. 4. Fermentation. QU 55 P96406 2006]
I. Shukla, Abhinav. II. Etzel, Mark. III. Gadam, Shishir. IV. Series.
QP551.P693 2006
612.3'98--dc22 2006002480
Visit the Taylor & Francis Web site at
http://www.taylorandfrancis.com
Taylor & Francis Group and the CRC Press Web site at
is the Academic Division of Informa plc. http://www.crcpress.com
© 2007 by Taylor & Francis Group, LLC
BIOTECHNOLOGY AND BIOPROCESSING SERIES
1. Membrane Separations in Biotechnology, edited by
W. Courtney McGregor
2. Commercial Production of Monoclonal Antibodies: A Guide for
Scale-Up, edited by Sally S. Seaver
3. Handbook on Anaerobic Fermentations, edited by Larry E. Erickson
and Daniel Yee-Chak Fung
4. Fermentation Process Development of Industrial Organisms,
edited by Justin O. Neway
5. Yeast: Biotechnology and Biocatalysis, edited by Hubert Verachtert
and René De Mot
6. Sensors in Bioprocess Control, edited by John V. Twork
and Alexander M. Yacynych
7. Fundamentals of Protein Biotechnology, edited by Stanley Stein
8. Yeast Strain Selection, edited by Chandra J. Panchal
9. Separation Processes in Biotechnology, edited by Juan A. Asenjo
10. Large-Scale Mammalian Cell Culture Technology, edited by
Anthony S. Lubiniecki
11. Extractive Bioconversions, edited by Bo Mattiasson and Olle Holst
12. Purification and Analysis of Recombinant Proteins, edited by
Ramnath Seetharam and Satish K. Sharma
13. Drug Biotechnology Regulation: Scientific Basis and Practices,
edited by Yuan-yuan H. Chiu and John L. Gueriguian
14. Protein Immobilization: Fundamentals and Applications,
edited by Richard F. Taylor
15. Biosensor Principles and Applications, edited by Loï'efc J. Blum
and Pierre R. Coulet
16. Industrial Application of Immobilized Biocatalysts, edited by
Atsuo Tanaka, Tetsuya Tosa, and Takeshi Kobayashi
17. Insect Cell Culture Engineering, edited by Mattheus F. A. Goosen,
Andrew J. Daugulis, and Peter Faulkner
18. Protein Purification Process Engineering, edited by Roger G. Harrison
19. Recombinant Microbes for Industrial and Agricultural Applications,
edited by Yoshikatsu Murooka and Tadayuki Imanaka
20. Cell Adhesion: Fundamentals and Biotechnological Applications,
edited by Martin A. Hjortso and Joseph W. Roos
21. Bioreactor System Design, edited by Juan A. Asenjo
and José C. Merchuk
22. Gene Expression in Recombinant Microorganisms, edited by
Alan Smith
23. Interfacial Phenomena and Bioproducts, edited by John L. Brash
and Peter W. Wojciechowski
24. Metabolic Engineering, edited by Sang Yup Lee
and Eleftherios T. Papoutsakis
25. Biopharmaceutical Process Validation, edited by Gail Sofer
and Dane W. Zabriskie
26. Membrane Separations in Biotechnology: Second Edition,
Revised and Expanded, edited by William K. Wang
© 2007 by Taylor & Francis Group, LLC
27. Isolation and Purification of Proteins, edited by Rajni Hatti-Kaul
and Bo Mattiasson
28. Biotransformation and Bioprocesses, Mukesh Doble,
Anil Kumar Kruthiventi, and Vilas Gajanan Gaikar
29. Process Validation in Manufacturing of Biopharmaceuticals:
Guidelines, Current Practices, and Industrial Case Studies,
edited by Anurag Singh Rathore and Gail Sofer
30. Cell Culture Technology for Pharmaceutical and Cell-Based Therapies,
edited by Sadettin S. Ozturk and Wei-Shou Hu
31. Process Scale Bioseparations for the Biopharmaceutical Industry,
edited by Abhinav A. Shukla, Mark R. Etzel, and Shishir Gadam
© 2007 by Taylor & Francis Group, LLC
Preface
Rapidgrowthinthebiopharmaceuticalindustryinrecentyears––bothinthe
valueandvarietyofnovelproducts—-hasturnedthisoncefledglingindustry
intoanimportantplayerintheglobaleconomy.Commercialsuccessofthese
productshingesonthesuccessfuldevelopmentandimplementationofrobust,
reliable, and economical production processes. Increasingly within the bio-
pharmaceutical industry, bioprocess development is seen as a key source of
competitiveadvantage.Thistrendwillcontinueastheindustryfurthermatures.
Bioseparations (also often called downstream processing) refers to the wide
varietyandcombinationsofproductionprocessesthatareemployedtorecover
and purify biomolecules from biological sources. Given the diversity of bio-
moleculesandthecomplexnatureoftheirbiochemicalpropertiesitisnowonder
thatthefieldofbioseparationshasevolvedintoarichandvariedone.
However, mostofthedevelopmentsinthebioseparationsfieldarechron-
icled in a rather vast range of scientific papers, patents, and conference
presentations that can pose a bewildering array for a newcomer to process
development or for established scientists and engineers who are seeking to
learnanunfamiliartechnique.Biologicalsystemsareinherentlycomplexand
veryoftencannotbedefinedpreciselybymathematicalmodels. Duetothese
reasons,bioprocessdevelopmentisoftensimultaneouslyanartandascience.
Additionally, biopharmaceutical manufacturing is a highly regulated activity
andhenceseveralregulatoryconsiderationsoftenenterintochoicesmadedur-
ing process development. This book first lays a foundation of basic concepts
andfundamentalprinciplesthatareessentialforunderstandingeachtopic,and
then provides a set of rules of thumb that are based on hands-on industrial
experiencewithactuallarge-scaleprocesses,hindsightlearningfromscale-up
problems, and regulatory issues that arise during development and licensure.
Thebookfollowsaconciseandpracticalapproachandisrepletewithtables,
flowcharts, andschematicsthatprovideaperspectiveonhowprocessdevel-
opment is carried out in the biopharmaceutical industry. In each chapter, the
authorsattempttobringtogetherscientificprinciples,practicalconsiderations,
and empirical approaches that are closely intertwined in this field to give the
readeraperspectiveonhowpurificationprocessdevelopmentisactuallycarried
outinthebiopharmaceuticalindustry.Inthissense,thebookisadeparturefrom
previoustextsinthisareathatprovideeitheralargelytheoreticalperspective
© 2007 by Taylor & Francis Group, LLC
SHUKLA: “dk3347_c000” — 2006/6/2 — 09:38 — page vii — #7
intothefieldorareacompilationofreviewpapersofscientificdevelopments
inanygiventopic.
Thebookcontainsthreebroadareasoffocus.Firstisafocusondownstream
unit operations, their fundamental principles, and considerations for process
development.Thisincludeschaptersonunitoperationsthatarewidelyaccepted
inthebioprocessingworldaswellasonmethodologiesthatcouldfindwider
acceptanceintheyearstocome.Nextcomesafocusonsomehighlyessential
ancillaryaspectsofdownstreamprocessdevelopmentincludingviralvalidation
and in-process analytical methods. The final chapters in the book deal with
downstreamprocessdevelopmentforvariousclassesofbiomoleculesandthe
strategiesadoptedfortheirprocess.
Chapter 1 presents a broad review of the principles of harvest clarifica-
tiontechnologies(centrifugation,depthfiltration,andtangentialflowfiltration)
alongwithacasestudyofharvestingatherapeuticproteinproductfromhigh
cell density fermentation broth. The comprehensive literature review within
this chapter should prove to be a valuable road-map for practitioners to nav-
igate this vastly studied area of downstream processing. Chapter 2 presents
theoretical and experimental frameworks and a real-life case study for devel-
opmentofexpandedbedadsorptionasanalternativetothemoreconventional
techniquespresentedinChapter1. Giventheincreasinginterestinthisinteg-
rativetechnologyinthelastdecadeanditspotentialtoreduceoverallcostof
goods, we believe this chapter will be a valuable resource for many readers.
Chapter 3 presents another novel technology (High Gradient Magnetic Fish-
ing) that can allow integration of harvest-clarification with chromatographic
captureandpurification.Magneticadsorbentshaveaverypowerfulandunique
“hook”thatpermitsoneto“fish”themfromcrudecellcultureandfermenta-
tionbrothssimplythroughtheapplicationofamagneticfield. Becauseofits
potentialforrapidprocessingoflargevolumesofcellharvest,thefutureindus-
trialprospectsforHighGradientMagneticFishinglookbright.Andthat’sno
fishstory.
Chapter 4 spells out the fundamental principles of protein refolding and
provides the reader with experimental strategies to develop and optimize a
refoldingprocess.Avarietyofusefulpointstoconsiderduringdevelopmentof
large-scaleproteinrefoldingoperationsareprovidedthroughoutthechapter.
Chapter 5 is on bulk protein crystallization –– a technique that is gener-
atingincreasinginterestfor(1)earlystagerecovery,(2)generatingultra-high
purity product in the polishing stages of a downstream process, (3) improv-
ingproductstabilityandshelflife, and(4)providingnewdosageformatsfor
proteintherapeutics.Thechaptersetsthestagefornewresearchersbydescrib-
ing the basic principles and key equations needed for design of experiments,
illustrating methods of data analysis, and providing case studies of industrial
practice.
© 2007 by Taylor & Francis Group, LLC
SHUKLA: “dk3347_c000” — 2006/6/2 — 09:38 — page viii — #8
Chromatographicseparationsareubiquitousinthemanufactureofbiophar-
maceuticals. For this reason, the next three chapters provide a tutorial, some
hands-onadvice,andagazeintothecrystalballforprocesschromatography.
Chapter 6 focuses on the different modes of chromatography. All the major
modesofprocesschromatographyarediscussed:affinity,ionexchange,hydro-
phobicinteraction,reversedphase,hydroxyapatite,immobilizedmetalaffinity,
thiophilic interaction, mixed mode, and size exclusion. Rules of thumb and
heuristicsbasedonlarge-scaleexperienceareprovidedineachsectiontoserve
asapracticalguideforthereader.
Chapter7presentspracticalconsiderationsandmethodologiesforscreen-
ing and selecting chromatographic resins for industrial separation processes.
Ofspecialmentionaretoolssuchashighthroughputscreening,retentatechro-
matography,andcumulativeyield–impurityplotsthatgreatlyfacilitatethetask
ofstationaryphaseselection.
Chapter8onaprioripredictionofchromatographicseparationsfrompro-
tein structure data describes a technique that might radically alter the current
paradigmofprocessdevelopment. Whiletheprecedingtwochaptersconcen-
trated on currently applied empirical methods for defining chromatographic
unit operations, this chapter describes some major strides toward the “holy
grail”ofresearchersinbeingabletopredictchromatographicperformanceby
simulationalone.Thechapterdescribesmethodologiesforpredictingchroma-
tographicparametersofproteinsandpresentsanoverviewofrecentadvances
madeinextendingthesepredictivetechniquesbeyondthesmallmoleculerealm
forwhichtheyhavebeenemployedsofar.
Thenextthreechaptersdealwithmembrane-basedunitoperations.Chapter
9presentssimplemathematicalmodelsforpredictingbreakthroughcurvesin
membrane chromatography systems, and explains how to use these models
to analyze laboratory and large-scale data. This analysis is extended to the
predictionofviralclearanceinmembranechromatographicsystems. Chapter
10 focuses on the design and implementation of an ultrafiltration step in
an industrial scale process. Ultrafiltration/diafiltration is ubiquitous in down-
stream processing for concentration, buffer exchange, and final formulation.
Special attention is paid to addressing engineering constraints faced during
scale-up of ultrafiltration systems. Chapter 11 —- on virus filtration process
design and implementation –– provides an overview of viral filter selection,
process design and optimization (in both normal and tangential flow mode),
and a detailed overview of the actual operating procedures for-full scale
implementation.
Recognizingtheeconomicpotentialoftransgenicsourcesforbiopharma-
ceuticalproduction,thebookincludesachapteronrecoveryofproteinsfrom
emergingtransgenicsources.Chapter12providesacomprehensiveeducational
overviewofthefieldofproteinproductionandrecoveryfromtransgenicsources
© 2007 by Taylor & Francis Group, LLC
SHUKLA: “dk3347_c000” — 2006/6/2 — 09:38 — page ix — #9
and also discusses the challenges that need to be addressed for transgenic
sourcedproductstobecomearealityinthefuture.
The next several chapters in the book deal with additional efforts aimed
at ensuring purity, efficacy, and safety in the development of downstream
processes for biopharmaceutical manufacturing. Key amongst these are in-
processanalyticalmethods,thatformtheeyesandearsofdownstreamprocess
developers.Chapter13dealswithvariousqualityandefficacyattributesofbio-
pharmaceuticalsandprovidespracticalguidanceonanalyticalmethodsthatcan
beemployedtoassessthem. Chapter14providesanexcellenttutorialonthe
elementsthatneedtobeinplaceinadownstreamprocesstoensuresafetyfrom
viralcontamination.Thechapterprovidesanoverviewofthevariouspotential
meansofviralintroductionintotheprocessstream,designofappropriatevirus
clearance studies, considerations in selecting appropriate model viruses, and
design of scale-down models. Chapter 15 describes the latest trends in viral
clearance,aregulatoryperspectiveofthisarea,andanin-depthcomparisonof
thevariousestablishedandupcomingmethodsofachievingviralclearance.
The final set of chapters in the book focus on various classes of bio-
moleculesandprovideinsightintotheirprocessscalepurification.Monoclonal
antibodies have emerged as one of the most important classes of biopharma-
ceuticals today and their downstream processing aspects are covered in three
chapters. Chapter 16 provides a comprehensive introduction to this class of
therapeuticsandprovidesdetailedpracticalguidancefordevelopingProteinA
chromatographyasthekeypurificationstepinantibodydownstreamprocessing.
Chapter17describesthedevelopmentofpolishingchromatographicstepsfor
monoclonalantibodydownstreamprocessing. Inadditiontoprovidinguseful
practical advice, the chapter also provides several useful process templates.
Purificationoflargebiomoleculessuchasgenetherapyvectorspresentsigni-
ficant challenges during scale-up when conventional chromatographic resins
areusedduetotheirlowbindingcapacities.Chapter18providesanindustrial
case-studyforanapprovedbiopharmaceutical(Remicade®)dealingwithpost
licensureprocesschangesandstrategiesemployedfortheirregulatoryapproval.
Chapter19describesthepurificationofabacterialpolysaccharidevaccine.
Investigation of an unexpected problem during scale-up of ultrafiltration for
thismoleculeledtoatroubleshootinginvestigationthatledtoabetterprocess
understanding.
Chapter 20 further highlights the advantages of convective transport in
membrane chromatography in overcoming capacity limitations in conven-
tionalbeadedchromatographicresinsforlargerbiomolecules.Thechapteralso
providesacomprehensiveliteraturereviewforthepurificationofgenetherapy
vectors—-anotheremergingclassofbiopharmaceuticals.
WehopeProcessScaleBioseparationsfortheBiopharmaceuticalIndustry
will be a valuable text for the growing numbers of scientific staff involved
© 2007 by Taylor & Francis Group, LLC
SHUKLA: “dk3347_c000” — 2006/6/2 — 09:38 — page x — #10
inprocessdevelopmentinthebiopharmaceuticalandbiotechnologyindustries,
academia,andgovernmentlaboratories.Insomeways,thisbookwasmotivated
bythesharedfeelingamongtheeditorsandauthorsthattherewasneedfora
comprehensive tutorial text combining fundamental principles and empirical
guidelinesoriginatingfromlarge-scaleexperienceinthebioseparationsarena.
Thismakesusconfidentthatthiswillalsobeatimelybookforgraduatestudents
andseniorlevelundergraduateswhoarepreparingforacareerinbioprocessing.
We believe this book will find a worldwide audience in the rapidly growing
biopharmaceuticalsector.
AbhinavA.Shukla
MarkR.Etzel
ShishirGadam
© 2007 by Taylor & Francis Group, LLC
SHUKLA: “dk3347_c000” — 2006/6/2 — 09:38 — page xi — #11
Editors
AbhinavA.ShuklaisPrincipalScientistinPurificationProcessDevelopment
atAmgenInc.inSeattle,Washington.Hisgroupisresponsibleforthedevel-
opment,characterization,validation,andtransferofdownstreamprocessesfor
bothearly-andlate-stagebiopharmaceuticals.Dr.Shuklahasbeeninstrumental
in setting up Amgen’s platform strategy for the purification of monoclonal
antibody therapeutics. He has also started several technology initiatives at
Amgen, and authored numerous publications and presentations in biosepara-
tions.PriortojoiningAmgenin2000,Dr.Shuklaheldasimilarroleinprocess
development at ICOS Corporation, Bothell, Washington dealing with down-
stream processing of bacterial and mammalian cell-culture-derived products.
He received his Ph.D. in chemical engineering from Rensselaer Polytechnic
Institute, Troy, New York and his B.S. and M.S. in biochemical engineering
andbiotechnologyfromtheIndianInstituteofTechnology.
Mark R. Etzel is Professor of Chemical and Biological Engineering at the
University of Wisconsin-Madison. Dr. Etzel has seventeen years of teaching,
research,andconsultingexperienceinbiologicalseparationprocessesinclud-
ing membrane adsorption and filtration, freeze drying and spray drying, ion
exchange and affinity chromatography, and protein crystallization. Professor
EtzelreceivedaB.S.atPurdueUniversity,andaPh.D.attheUniversityofCali-
forniaatBerkeley, bothinthedepartmentofchemicalengineering. Dr.Etzel
workedinindustryforsixyearsbeforereturningtoacademia.
Shishir Gadam is part of the Bioprocess Research and Development divi-
sionatMerckandCo., Inc., WestPoint, Pennsylvania, andisresponsiblefor
clinicalmanufacturingofbulkvaccineandtherapeuticproteinsusingMerck’s
state-of-the-artmultiproductbiologicspilotplant.Duringhislastnineyearsof
stay at Merck he has contributed in various areas within biologics including
process development, scale up, technology transfer to manufacturing, GMP
manufacturingforclinicaluse,andprocessvalidation.PriortojoiningMerck
in 1997, Dr. Gadam spent three years at Millipore Corporation, Bedford,
Massachusetts, developing new and novel membrane purification technolo-
gies. Dr.GadamobtainedhisPh.D.inchemicalengineeringfromRensselaer
PolytechnicInstitute,Troy,NewYork.
© 2007 by Taylor & Francis Group, LLC
SHUKLA: “dk3347_c000” — 2006/6/2 — 09:38 — page xiii — #13