Therapeutic Antibodies: Methods and Protocols

586 Pages·2009·5.64 MB·English

by  Dimiter S. Dimitrov

Therapeutic Antibodies
M E T H O D S I N M O L E C U L A R B I O L O G Y TM
John M. Walker, SERIES EDITOR
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M M B TM
E T H O D S I N O L E C U L A R I O L O G Y
Therapeutic Antibodies
Methods and Protocols
Edited by
Antony S. Dimitrov
Profectus BioSciences Inc.
Baltimore, MD, USA
Editor
AntonyS.Dimitrov
ProfectusBioSciencesInc.
6411BeckleyStreet,
Baltimore,MD21224
USA
[email protected]
SeriesEditor
JohnM.Walker
UniversityofHertfordshire
Hatfield,Herts.
UK
ISSN1064-3745 e-ISSN1940-6029
ISBN978-1-934115-92-3 e-ISBN978-1-59745-554-1
DOI10.1007/978-1-59745-554-1
LibraryofCongressControlNumber:2008942048
#HumanaPress,apartofSpringerScienceþBusinessMedia,LLC2009
Allrightsreserved.Thisworkmaynotbetranslatedorcopiedinwholeorinpartwithoutthewrittenpermissionofthe
publisher(HumanaPress,c/oSpringerScience+BusinessMedia,LLC,233SpringStreet,NewYork,NY10013,USA),
except for brief excerpts in connection with reviews or scholarly analysis. Use in connection with any form of
informationstorageandretrieval,electronicadaptation,computersoftware,orbysimilarordissimilarmethodology
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Printedonacid-freepaper
springer.com
To My Parents Maria and Stancho,
and Brother Dimiter
Preface
Over 2000 years ago in China, antibodies elicited by early forms of vaccination likely
playedamajorroleintheprotectionofthepopulationfrominfectiousagents.Vacci-
nation has been further developed in Europe and described by Edward Jenner in the
late-eighteenth century, then successfully implemented worldwide. The idea to use
theactiveingredientinthebloodofvaccinated(orimmunized)animalsorhumansfor
the treatment of diseases came a century later. It was made possible by a series of
discoveries,suchastherealizationthattheserumfromanimalsimmunizedwithtoxins,
for example, diphtheria toxin or viruses, is an effective therapeutic against the disease
causedbythesameagentinhumans.Inthe1880s,vonBehringdevelopedanantitoxin
(anti-body) that did not kill the bacteria but neutralized the bacterial toxin. The first
Nobel Prize in Medicine (1901) was given to him for the discovery of the serum
therapy.Acenturylater,22monoclonalantibodies(mAbs)areapprovedbytheUnited
States Food and Drug Administration (FDA) for clinical use, and hundreds are in
clinicaltrialsforthetreatmentofvariousdiseasesincludingcancers,immunedisorders,
and infections. The revenues from the top-five therapeutic antibodies reached $11.7
billion in 2006, and major pharmaceutical companies raced to acquire antibody
biotech companies with a recent example of MedImmune, Inc., which was acquired
for$15.6billionbyAstraZenecain2007.
This explosion of research and development in the field of therapeutic antibodies
prompted the publication of the MiMB volume Therapeutic Antibodies: Methods and
Protocols. The book’s major goal is to present a set of protocols useful for researchers
discoveringanddevelopingtherapeuticantibodies.Currentadvancesandfuturetrends
in the antibody therapeutics are analyzed in the lead-in review article. The road from
identificationorselectionofappropriatetargetstoantibodiesinclinicaluseisdivided
into five major stages: (1) recombinant antigens, (2) antibody libraries, (3) antibody
discovery,(4)antibodyengineering,and(5)antibodypreclinicaldevelopment.Alsoa
low-cost antibody sequence database is described in the last chapter. Representative
protocolsforeachstagearewrittenbyleadingexpertsfromacademiclaboratoriesand
biotechnology companies. Protocols for antibodies as reagents are not included
becauseof theexistence ofexcellent books onmethods for such antibody generation
andcharacterization.
PartIincludesseveral methodsthathavebeensuccessfullyemployedtoproduce,
purify,andcharacterizesolublesecretedversionsofseveralviralenvelopeglycoproteins
successfully used as antigens for selection of neutralizing human monoclonal antibo-
dies.PartIIdetailsmethodstocreatephagelibrariesofhumansyntheticsingle-chain
antibodies, human antibody domains (V ), and rabbit antibodies. It also details a
H
method for construction of a large na¨ıve human Fab library, which was successfully
usedforselection ofpotent neutralizingantibodiesagainstviruses andcancer-related
proteins. Part III contains protocols for selecting antibodies against intracellular
targets,specificinternalizationfragments,antibodieswithbroadspectrumofbinding
vii
viii Preface
and neutralization, non-aggregating V binders from synthetic phage libraries, and
H
IgGs from combinatorial libraries expressed in Escherichia coli. It also contains
advancedmethodsforhigh-throughputscreeningofsingle-chainantibodies,identifi-
cationoffullyhumanantigen-specificantibodyrepertoirefromplasmacells,andrapid
screeningplatformforstabilizationofsingle-chainantibodies.PartIVcoversmethods
for antibody engineering including affinity maturations, construction of tetravalent
bispecific antibodies, deimmunization of antibodies, and preparation and character-
ization of antibody conjugates for targeted cancer therapies. Part V describes several
aspects of the antibody preclinical development including high-level production for
laboratory studies, scaling up and production for preclinical animal studies, in vitro
antibody potency and breadth of virus neutralization, and in vivo methods for estab-
lishingsynergybetweenantibodiesincancertherapyinmiceandpassiveimmunization
againstHIV-1inmacaques.
I am indebted to all contributing authors for sharing their expertise, to Professor
JohnM.WalkerforinvitingmetoeditthisvolumeofMiMBseries,andtoProfectus
BioSciences, Inc. for their support during the preparation of the book. Finally, I am
gratefultomychildrenMilenaandStanislav,whohaveencouragedmeintheventure
ofeditingthisbook.
AntonyS.Dimitrov
Contents
Preface.................................................................vii
Contributors............................................................ xiii
ColorPlates ............................................................xvii
1 TherapeuticAntibodies:CurrentStateandFutureTrends–IsaParadigm
ChangeComingSoon? .................................................1
DimiterS.DimitrovandJamesD.Marks
PART I: RECOMBINANT ANTIGENS
2 PreparationofRecombinantViralGlycoproteinsforNovelandTherapeutic
AntibodyDiscovery ...................................................31
Yee-PengChan,LianyingYan,Yan-RuFeng,andChristopherC.Broder
PART II: ANTIBODY LIBRARIES
3 DesignofaHumanSyntheticCombinatorialLibraryofSingle-Chain
Antibodies ..........................................................61
LimorNaharyandItaiBenhar
4 ConstructionofaHumanAntibodyDomain(V )Library.....................81
H
WeizaoChen,ZhongyuZhu,XiaodongXiao,andDimiterS.Dimitrov
5 GenerationandSelectionofRabbitAntibodyLibrariesbyPhageDisplay.........101
ChristophRader
6 ConstructionofaLargeNa¨ıveHumanPhage-DisplayedFabLibrary
ThroughOne-StepCloning............................................129
ZhongyuZhuandDimiterS.Dimitrov
PART III: ANTIBODY DISCOVERY
7 IdentificationofTargetandFunctionSpecificAntibodiesforEffective
DrugDelivery ......................................................145
YuZhouandJamesD.Marks
8 ScreeningofSpecificInternalizationFabFragmentfromHumanNaive
PhageLibrarybyCombinationalBio-Panning..............................161
XinWangandBrianB.Cao
9 CompetitiveAntigenPanningforSelectionofHIV-1NeutralizingHuman
MonoclonalAntibodiesSpecificforgp41 .................................175
Mei-YunZhangandDimiterS.Dimitrov
10 SelectionofNon-aggregatingV BindersfromSyntheticV Phage-Display
H H
Libraries...........................................................187
MehdiArbabi-Ghahroudi,RogerMacKenzie,andJamshidTanha
ix
x Contents
11 IsolationofFull-LengthIgGAntibodiesfromCombinatorialLibraries
ExpressedinEscherichiacoli............................................217
YarivMazor,ThomasVanBlarcom,BrentL.Iverson
andGeorgeGeorgiou
12 MultiplexedFlowCytometry:High-ThroughputScreeningofSingle-Chain
Antibodies .........................................................241
JoanneAyriss,RosaValero,AndrewR.M.Bradbury,andPeterPavlik
13 HumanAntibodyRepertoires ..........................................261
Per-JohanMeijer,LarsS.Nielsen,JohanLantto,andAllanJensen
14 RapidScreeningPlatformforStabilizationofscFvsinEscherichiacoli............279
BrianR.Miller,ScottM.Glaser,andStephenJ.Demarest
PART IV: ANTIBODY ENGINEERING
15 InVitroAntibodyAffinityMaturationTargetingGermlineHotspots............293
MitchellHoandIraPastan
16 AffinityMaturationbyPhageDisplay ....................................309
HolgerThie,BerndVoedisch,StefanDu¨bel,MichaelHust,
andThomasSchirrmann
17 ProductionofChimericHeavy-ChainAntibodies...........................323
JianbingZhang,RogerMacKenzie,andYvesDurocher
18 MammalianCellDisplayforAntibodyEngineering .........................337
MitchellHoandIraPastan
19 ImprovingAntibodyBindingAffinityandSpecificityforTherapeutic
Development.......................................................353
JennyBostrom,ChingweiV.Lee,LauricHaber,andGermaineFuh
20 ConstructionandProductionofanIgG-LikeTetravalentBispecific
AntibodyforEnhancedTherapeuticEfficacy...............................377
DanLuandZhenpingZhu
21 DeimmunizationofMonoclonalAntibodies ...............................405
TimD.Jones,LauraJ.Crompton,FrankJ.Carr,andMatthewP.Baker
22 Anti-CD22Onconase:PreparationandCharacterization .....................425
DianneL.Newton,LukeH.Stockwin,andSusannaM.Rybak
23 Antibody–CytotoxicAgentConjugates:PreparationandCharacterization........445
RajeevaSinghandHansK.Erickson
PART V: ANTIBODY PRECLINICAL DEVELOPMENT
24 High-LevelProductionofaHumanizedImmunoRNaseFusionProtein
fromStablyTransfectedMyelomaCells...................................471
Ju¨rgenKrauss,EvelynExner,AthanasiosMavratzas,SiegfriedSeeber,
andMichaelaA.E.Arndt
25 AntibodyFragmentExpressionandPurification ............................491
DimanaDimitrova,ViditaChoudhry,andChristopherC.Broder
26 Scaling-UpandProductionofTherapeuticAntibodiesforPreclinical
Studies............................................................499
YangFengandDimiterS.Dimitrov