MAbs.
`DUP — General Collection
`
`W1 MA144C
`
`v. 1 , no. 4
`JLII<Aug 2009
`
`
`
`J
`Volume 1 0 Issue 4 0 July/August 2009
`
`Editor-in-(hief
`Janice M. Reichert
`
`,/
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`
`
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`%i§ NATIONAL
`.
`LIBRARY OF
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`MEDICINE
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`PROPERTY OF THE
`
`348 light-Activatable Bispetific (oniugutes
`
`370 Improving Antibody Production
`
`BIOSC I [ENCE
`
`382 Patenting Antibodies
`
` 332 Allotypes and immunogenicity
`
`www.Iondesbioscience.tom/iournuls/mabs
`
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`Edirorial 0mm:
`
`Editor-in-Chief
`
`Lamlcs liioscicncc
`1002 Won Avenue. 2nd liloor
`Armin. Toms 78701 USA
`5l2,6_37.(1050 phone
`512,657,607‘) lint
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`Journal Publications Director
`Kimberly A Mitchell
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`Managing Editor
`Nicolc’lbiltl
`nicoch‘l.intlcshioscicncc,c0m
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`Arr Direction/Production Director
`K.“ h ryn Sauccda
`knKG‘lanrlcsl)ioscicncuom
`
`Janice M. Reichcrt
`Tufts Centerfor flat Study ofDrug Development
`Boston, Massachusetts USA
`
`Editorial Board
`
`Gregory P. Adams
`Fox Chase Cancer Center
`
`Philadelphia, PA USA
`S. Robcrr Adamson
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`“filer/I Iiiolt'r/I
`AIM/iron, N] USA
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`iWiIJ/o C/infr Tram/Jan! Center
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`Shite UIIiI/clsity ofNeu/ York
`at [it/flo/o
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`Centre 41Immunologic I’ierre
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`Seattle, WA USA
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`Michcl Chm-train
`[Merck ("1" Co. Inc.
`Rir/ony, N] USA
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`University ofDe/in' Sum/1
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`New York, NY USA
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`PI)L Biop/nznn/z
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`chcor, Inc.
`Alonrooin, CA USA
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`NCI. NIH
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`Ar/Ilent Venture Partners
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`7Ec/1nim/ Unix/mi1y affirmin—
`rr/Iwcig
`Brmmo'lmaeig, Germany
`Martin Glennie
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`Unit/mil] ofSout/nInI/Iton
`Som/nunplon, UK
`David Glover
`Air/[Cell Iiioscience. Llri‘
`Gunilridgc, UK
`lqbal S. Grcwal
`Sam/e Genetics. Inc.
`Bar/1c”. WA USA
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`Thi: mafaxial “a: rrknind‘l
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`Yajun Guo
`Second Ali/nor} Alm'im/
`University
`S/nzng/nzi, C/n'rnr
`Sherif Hanala
`Clo/nil BioFocm LLP
`San I“ran(irro, CA USA
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`NCI. NIH
`Bet/Mild. All) USA
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`P. Mark Hogarth
`Burner Institute
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`Hennic R. Hooge‘nboom
`Aowa
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`James S. Huston
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`University ofBirming/Irml
`Birmingbmn, UK
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`Cenlomr RU'D. Inc.
`Roi/nor; PA USA
`Gnurav Laroia
`Alcrc/r 6? C0,, Inc.
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`IMGI; CNRS, Uniocrsilt‘
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`[Her/are):
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`Mil/aims, CA USA
`Henry Lowman
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`Soot/1 San I‘rzlncirco. CA USA
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`James D. Marks
`University oszliforni/I,
`San Francisco
`San Francisco, CA USA
`
`John L. Marquardt, Jr.
`Pinnegrm. Henderson, For/zoom,
`Gormt C'TDunner, LIP
`W/rtr/Jinglon DC USA
`RA. Masllelkar
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`Nation/II CI/emiml Liz/lorrltoljr
`I’nne, Inziizz
`Richard Mason
`BTG Ltd,
`London UK
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`John McCafl'erty
`University ofCambridge
`Cmn/Irige UK
`Jan ter Meulcn
`[Merck Rere/Irc/J Labor/norm
`“75! Point. PA USA
`Sheri: Morrison
`
`University oszz/iforni/z. L05
`Ange/es
`LosAngc/ex, CA USA
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`Longrn, Germany
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`Swiss 1%ch Institute
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`ofYET/1710100
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`Jane Osbourn
`McriImmunc, Inc.
`Cambridge, UK
`Gilles Paintaud
`
`Francois Robe/air Uniocrrity
`and CNRS
`Toxin; I’mnce
`Ira I’astan
`NCI. NIH
`Bet/Mylo, MD USA
`
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`
`
`Volume] ' Issue4 . July/August 2009 ' 307-392
`
`CONTENTS
`
`Editorial
`
`
`307
`
`Letter from the Editor
`
`Janice M. Reichert
`
`Meeting Reports
`
`
`308
`
`3'8
`
`5"“ Annual Monoclonal Antibodies Conference:
`March 24—25, 2009, London, UK
`
`Mari Herigstad and Lisa Urquhart
`
`Keystone Symposium on Antibodies as Drugs:
`March 27—April 1, 2009, \Whistler, BC CA
`Thierry Wurch, Christel Larboutet and Bruno Robert
`
`M i n i - R e v i e w
`
`326
`
`Ofatumumab
`
`Bodi Zhang
`
`R e v i e w
`
`
`332
`
`Human immunoglobulin allotypes: Possible implications
`for immunogenicity
`Roy Jeffetis and Marie—Paulo Lefranc
`
`R e p o r t s
`
`
`339
`
`348
`
`Molecular construction and optimization of anti-
`human lL-lOt/B dual variable domain immunoglobulin
`(DVD-Igm) molecules
`Chengbin Wu,
`l-lua Ying, Sahana Bose, Renee Miller,
`Limaty Medina, Ling Santora and Tariq Ghayur
`
`Preclinical evaluation of light-activatable, bispecific anti-
`human CD3 antibody conjugates as anti-ovarian cancer
`therapeutics
`Stephen Thompson, John Dessi and Colin H. Self
`
`357
`
`Generation and characterization of chicken monoclonal
`
`antibodies against hutnan LOX-1
`Shin Iwamoto, Norihisa Nishimichi, Yoshiko Tateishi,
`
`Yuko Sato, Hiroyuki Horiuchi, Shuichi Furusawa, Tatsuya
`Sawamura and Haruo Matsuda
`
`364
`
`370
`
`Identification of circulating neuropilin—l and dose—
`dependent elevation following anti—neuropilin-l anti-
`body administration
`Yanmei Lu, Hong Xiang, Peter Liu, Raymond R.
`Tong, Ryan J. Watts, Alexander W. Koch, Wendy N.
`Sandoval, Lisa A. Damico,Wai Lee Wong and Y. Gloria
`Meng
`
`Inhibitory effects of persistent apoptotic cells on
`monoclonal antibody production in vitro: Simple
`removal of non-viable cells improves antibody produc-
`tivity by hybridoma cells in culture
`Christopher D. Gregory, John D. Pound, Andrew
`Dcvitt, Megan Wilson—Jones, Patthasarathi Ray and
`Ruth J. Murray
`
`P o i n t of Vi e w
`
`
`397
`
`Non—human primate immune libraries combined
`with germline humanization: An (almost) new, and
`powerful approach for the isolation of therapeutic
`antibodies
`
`Thibrtut Pelat and Philippe Thullier
`
`Special Focus:
`Patenting Antibodies
`
`
`382
`
`385
`
`the gate: Patent protection for thera-
`Guardians at
`peutic monoclonal antibodies—Part 1
`Kevin W. McCabe
`
`Patenting antibodies in Europe
`Louise Holliday
`
`Editor's Corner
`
`
`387
`
`390
`
`Probabilities of success for antibody therapeutics
`Janice M. Reichett
`
`Upcoming meetings
`Janice M. Reichert
`
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`

`
`
`Volumei - Issue4 ° July/August 2009 - 307-392
`
`CONTENTS
`
`Deparhnenis
`
`39]
`
`Guidelines for Authors
`
`Aboutthe Cover
`
`
`
`
`Human diseases are complex and often involve multiple mediators con-
`tributing to disease pathogenesis, Therefore, a therapeutic agent that
`inhibits multiple disease mediators Will likely provide better. therapeutic
`efficacy. By genetically linking the variable domains of two dlSllnCl mego
`clonal antibodies in both heavy chain and light chain, a dual- spec; I:
`tetravalent lgG-like molecule, termed dual variable damaln ImmEmOQ'O
`ulin (DVD-lg), is generated with dual targeting capability, rnanulaclurlng
`leasibility and drug-like properties. The DVD-lg class oi mocfcu T55 repre-
`sents a new generation oi biologic agent For therapeutic
`eve opmen .
`For additional inlormation, see Wu et al., pp. 339—47.
`
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`

`[mAbs 1:4, 332-338,- July/August 2009]; ©2009 landes Biosdence
`
`Review
`
`Human immunoglobulin allotypes
`
`Possible implications for immunogenicity
`
`Roy Jefferis” and Marie-Paule Lefranc2
`
`ISchool of Immunity and Infection; College of Medical and Dental Sciences; University of Birmingham; Birmingham, UK; QIMGT; Laboratoire d’Immuno Généiique Moléculoire;
`Institut de Génétique Humaine; Montpeilier, France
`
`IgG
`imnnmogenicity, anti-therapeutic antibody,
`IgG allotypes, antibody therapeutics,
`Key words: human IgG, polymorphisms,
`glycosylation
`
`More than twenty recombinant monoclonal antibodies are
`approved as therapeutics. Almost all of these are based on the
`whole IgG isotype format, but vary in the origin of the variable
`regions between mouse (chimeric), humanized mouse and fully
`human sequences; all of those with whole IgG format employ
`human constant
`region sequences. Currently,
`the opposing
`merits of the four IgG subclasses are considered with respect to
`the in vivo biological activities considered to be appropriate to
`the disease indication being treated. Human heavy chain genes
`also exhibit extensive structural polymorphism(s) and, being
`closely linked, are inherited as a haplotype. Polymorphisms
`(allotypes) within the IgG isotype were originally discovered
`and described using serological reagents derived from humans;
`demonstrating that allotypic variants can be immunogenic and
`provoke antibody responses as a result of alIo—immunization. The
`serologically defined allotypes differ widely within and between
`population groups; therefore, a mAb of a given allotype will,
`inevitably, be delivered to a cohort of patients homozygous for
`the alternative allotype. This publication reviews the serologi-
`calIy defined human IgG allotypes and considers the potential
`for allotype differences to contribute to or potentiate immuno—
`genicity.
`
`Introduction
`
`Recombinant human protein (r?) and glycoprotein (tGP)
`therapeutics are established in the clinic. However, a variety of
`adverse reactions are reported that may differ between individual
`therapeutics, between the saute therapeutic produced by different
`companies or different lots produced by the same company. Even
`when these parameters are controlled, there still remains the final
`“black box:” the patient and individual disease manifestations. A
`
`"Correspondence to: Roy Jefferis; School of Immunity and Infection; College of
`Medical and Dental Sciences; University of Birmingham; Edgbaston, Birmingham
`BI5 211' UK; Email: r.ieerris@bham.ac,uIt
`
`Submitted; 05/05/09; Accepted: 05/28/09
`
`Previously published online as a mAbs E-publication:
`www.|andesbioscience.com/journais/mabs/article/9] 22
`
`feature of all rI’/rGI’ is a potential to be immunogenic i.e., cause
`the generation of anti-therapeutic antibodies (ATA). Such anti—
`bodies may neutralize the therapeutic, result in enhanced clearance
`or precipitate severe adverse reactions. To limit the generation of
`ATA, an rP/rGI’ should, ideally, have exactly the same structure as
`the natural product since departure from such structural fidelity
`constitutes “altered-self," and the potential to be immunogenic.
`The defined molecular structure of a natural protein/glyco-
`protein results from multiple inrra-cellular processes that include
`co- and post-translational modifications (CTM; I’TM). However,
`it
`is
`important
`to recognize that
`the structure assigned to a
`“natural" protein/glycoprotein is determined for molecules that
`have been resident in bodily fluid(s), prior to isolation and purifi—
`cation employing multiple protocols. In contrast, human rI’/rGI’
`therapeutics are produced in xenogcneic tissues such as Chinese
`hamster ovary (CHO cells) or mouse NSO cells that may yield
`product without the necessary human type CTM and I’TM, or
`add non—human CTM and I’TM. Following secretion, the rI’/rGI’
`is exposed to the culture medium,
`to products of the producer
`cell line and subject to rigorous down—stream processing, formula-
`tion, storage and a defined delivery protocol. High productivity
`may compromise the cellular machinery for CTM and I’TM
`with consequent poor product quality; it
`is essential therefore to
`apply qualitative as well as quantitative criteria at an early stage in
`clone selection. Despite these difficulties, both actual and poten—
`tial, rapid progress has been made for both the productivity and
`stability ofrP/rGP produced in mammalian cells. These issues have
`been explored in numerous review articlesdj‘
`Whilst the above parameters that might predispose antibodies
`to be immunogenic are widely recognized, and appropriate steps
`taken to monitor and control them, the consequences ofadmin—
`istration to diverse human populations is less chI developed.
`Human populations exhibit multiple genotypes and phenotypes,
`and one would not Wish to add structural differences due to
`
`polymorphisms to the inherent propensity for rI’erI’ products
`to exhibit structural heterogeneity. However, all currently licensed
`recombinant IgG antibody therapeutics have been developed as
`a single polymorphic (allotypic) form;
`therefore, administration
`to patients homozygous for the alternative allotype(s) presents
`
`332
`
`mAb:
`
`2009; Vol. I Issue 4
`
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`

`

`Human immunoglobulin allotype:
`
`added potential for immunogenicity. This review is intended to
`provide a summary of the serologically defined allotypes present
`within human IgG heavy and light chains, and the potential for
`recombinant monoclonal antibody therapeutics (mAbs)
`to be
`immunogenic when administered to diverse human populations.5
`
`Immunogenicity of mAbs
`
`is no surprise that Orthoclone-OKT3 (muromonab), a
`It
`licensed mouse lgGZa anti-CD3 antibody used in treatment of
`acute rejection in transplant patients, is immunogenic in humans,
`i.c.. causes development of anti—therapeutic antibody (ATA)
`responses in patiertts.("8 However, use ofthis drug continues as it
`is effective and the patient is only exposed to it for a short time.
`A significant reduction in itnmunogenicity and ATA was achieved
`with the development ofchimeric antibodies, bearing mouse heavy
`and light chain variable domains (VH, VL) and human constant
`regions (CII, CL). Humanization of the mouse VH and VL, by
`”transplanting," at the DNA level, the mouse or rat complemen—
`tarity regions (CDR)
`into human VI—I/VL framework regions
`(FR) further reduced potential itntnunogenieity. In practice, such
`“lunnanized” domains usually resulted in reduced affinity or speci—
`ficity such that sotne mouse or rat residues had to be re-introduced,
`with consequent
`increase in potential
`immunogenicity. With
`the advent of “fully" human antibodies, generated from phage
`display libraries or mice transgenic for human immunoglolmlin
`genes,
`it was anticipated that immunogenicity and the develop—
`ment ofATA might be circtlriiveritetl.6‘8 In practice the first such
`antibody, adalimumab (Hutnira) resulted in ~120/o incidence of
`ATA in patients with rheumatoid arthritis. This was subsequently
`reduced to ~2—5% for patients also receiving the non—steroid anti-
`inflammatory drug methotrexate, a mild itnmunosuppressant.9
`Initially, one may wonder why a fully human IgG mAb should
`be immunogenic; however, on reflection it may seem inevitable.
`The hallmark of an antibody is
`its specificity, not
`just
`for a
`particular target, but also for a unique structural feature on that
`target—the epitope. This is achieved within a secondary immune
`response that
`is characterised by somatic hypermntation and
`selection. Thus, each antibody is a structurally unique molecule
`with a unique epitope-binding site, the paratope. mAbs generated
`from phage display libraries or transgenic mice are unique to an
`individual, human or mouse, and may be perceived as "foreign"
`to a unique recipient. Given this inherent potential for immuno—
`genicity, other structural disparities should be avoided, Allotypic
`variations constitute such a disparity between individuals and
`population groups.
`
`Human IgG Isotypes, Allotypes and Idiotypes
`
`Human antibodies or imtnunoglobulins (Ig) were originally
`defined as gamtna (Y) globulins because of their mobility to the
`gamma region on paper electrophoresis. Subsequently, nine isotypes
`were defined structurally and serologically, mostly with antisera
`raised in guinea pigs or rabbits. The nine isotypes (iso = same)
`are present in normal human serum and each isotype expresses a
`unique profile ofeffector functions.”12 Each isotype was further
`defined by the unique amino acid sequence ofthe constant regions
`
`of their heavy chains and subsequently the sequence of the IGHC
`genes that encode these constant regionsda”14 Protein sequencing
`revealed the CH regions to be comprised ofthtee or four repeating
`homologous sequences (domains). This was confirmed at the gene
`level by the demonstration that each domain is encoded within
`a separate exon, interspersed by introns. The heavy chains of the
`classes are designated as 0L (IgA), p (IgM), 5 (IgD), Y (IgG), 8
`(lgE), respectively. The y heavy chains of the IgG comprise three
`domains, CH1, CH2 and CH3, with a “hinge” sequence inter—
`vening between the CHI and CHZ domains. When referring to
`individual IgG subclasses, the heavy chains are enumerated as Y1,
`Y2, Y3 and Y4, respectively. Two types of light chain, kappa (K)
`and lambda (7»), were also originally defined serologically, and
`subsequently by protein and gene sequences. Each H2L2 module
`expresses either two kappa or two lambda light chains to form
`H2K2 or H212 hetero—dimers.
`
`Allotypy within human IgG was first described by Grubb
`who showed that certain human sera would agglutinate erythro-
`cytes sensitized with human incomplete anti—Rh antibody.15‘l7
`Extensive polymorphism (allotypy) within human IgG heavy and
`light chains was subsequently recognized by careful serological
`typing, using human reagents obtained from tnultiparous women,
`multiple transfused individuals and normal blood donors (6%).
`Thus,
`the discovery of this polymorphism demonstrates that
`exposure of an individual to IgG ofa non-self allotype can induce
`an anti—allotype response. Gene sequencing studies have revealed
`extensive structural polymorphisms“14 however, this review will
`only consider serologically defined allotypes and their amino acid
`sequence correlates.
`By definition allotypes are shared amongst individuals within
`population groups, although studies in rabbits showed that anti-
`bodies raised within an individual rabbit could be immunogenic
`to another rabbit ofthe same allotype. The unique epitopes recog-
`nized were termed idiotypes. The term idiotype is now in common
`use to indicate the uniqueness of a monoclonal antibody, although
`definition of the term has lead to controversial debate and esoteric
`arguments in the past.
`It might be instructive,
`therefore,
`to
`consider its origin in more detail, Extensive allo—immumzanons
`and serological studies revealed and characterized serologically
`defined allotypy within rabbit heavy and light chains. By exten—
`sion, Oudin hyper—immunized rabbits with Salmonella typ/Jz' and
`used the sera to generate immune complexes that were adminis—
`tered to other rabbits of the same allotype. The recipient rabbits
`produced a serologically unique response that was specific for the
`antibodies raised in the donor rabbit and did not cross-react with
`anti—Salmonella typ/Jz' antibodies raised in other rabbits of the same
`allotype.” In a definitive publication, the terms isotype and allo-
`type were extended to include idiotype and justified as follows:18
`”T/Jefz'rrtprtrt oft/1e word (idio, from Greek, peculiar) it jztttz'fial [7y
`the extreme peculiarity of the antigenic tpcclficitiet in qrmtimz. An
`idiotype [5 a peculiar leiml ofpratein antigen defined by its izlintypic
`specificity. "
`The important point is that idiotypy was defined, setologically,
`following allo-imtnunization protocols. At the same time Kunkel
`was investigating human monoclonal proteins isolated from the
`
`www.lundcsblos¢lente.tom
`
`mill):
`
`333
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`

`

`‘l'li
`
`n(.'lml,2
`
`gem
`
`IamVIWLMa
`v V
`.1.—
`CH3
`V V
`—
`
`(y'lrnl.2
`
`DWIWGK; (.'m
`
`VL
`— ‘'
`(.‘lm3
`
`CL
`" "
`R _
`
`(ilml7
`
`CHI
`
`KznV
`
`'H A-l
`Km]
`Kmll .»\,.,._4.,.,,
`Kim?
`[\m-Vm
`
`Allotypy and Haplotypes Within Human lgG Heavy and
`Light Chains
`
`in 1976 the World Health Organization sponsored an expert
`committee meeting at which the nomenclature for lnrrnan immu—
`noglobulin allotypes was systemarizcd and a numerical system
`was proposed to replace the alphabetical system (Table llfu'l‘l
`Both systems may be encountered in the literature; particularly
`when reference is tirade to original publications in which the allo—
`types were defined. Allotypcs of lgG proteins are defined by the
`expression of unique epitopcfs)
`recognized by unique scrologic
`reagentfs). Allotypes expressed on the constant
`region of 1g(i
`heavy chain are designated as (im ((ieneric marker) together with
`the subclass, e.g., (11m, and the allotype number (or letter), erg.
`(Slml [or (.ilmia)], (Brni [or(1,5mfbl)]. Polymorphisms within
`lgA and kappa light chains are designated A2m, e.g.. Alml.
`and Km, e.g., Krill
`respectively. Serological polymorphisms have
`not been reported for lambda chains; hchver, there are multiple
`lambda chain isotypes and the rrnrnber oi' l(il.(3 ((11) genes can
`vary between individuals.“"3
`All the allotypes presented in 'l'able 1 are expressed on immuno—
`globulin constant regions. Since the genes encoding the constant
`region of the heavy chains (1(111C) are closely linked within
`the Kill gene locus they are inherited together as a haplotype
`with a low frequency of crossovers.
`llowcvcr, crossover events
`have occurred during evolution resulting in present populations
`expressing characteristic ltaplotypes‘, hence the usefulness of the
`allotype system in population studies, Table 2.1”“ Thus,
`in
`northern Europe (The Netherlands)
`(llc haplotype (irttS';5;2.’J
`that corresponds to the allotypcs
`(i3mS.|(],l],1_‘}.1/l,2(t,27;(i
`lrnj;(i2ml3 is present with a frequency of (MS and virtually
`all
`individuals are homolygons for the Alml allele. However,
`amongst Nigerian Africans the (imS';17;l haplotype occurs with
`a frequency ofiii.()78, the ( itrr5*;5;2.5 haplotype is not encountered
`and the AZmZ allele is present with a frequency oiifl.82(r. Prior to
`the development of. DNA fingerprinting techniques the distribu—
`tion of'Urn haplotypes between individuals and population groups
`was employed in population studies, paternity testing and forensic
`7.7-5]
`science’
`and provided a bridge betWeen immunological and
`molecular analysis 011“.“
`
`Allotype Expression and Structural Correlates With Amino
`Acid Residues (Eu/lmgt Numbering)
`
`lgGl. The heavy chains of lgGl proteins may express (.ilmj.
`Glml7,1 or
`(.ilml7.1,2 allotypes or
`(ilmil').
`(ilmfltsll,
`Glrrt(1,a,x), I‘espeClivcly.lt’ The structural correlations with amino
`acid residues are illustrated in Fig.
`I;
`thus the constant
`region
`of'GIml7,1, and Glm17,1,2 heavy chains differ from that of
`Glut} heavy chains by three and four arrrino acids, r'e.sl,)ectively,lS
`The presence oliC111 argininc (R) at position 214 (lMGil' R1 20;
`IMGT‘E’ www.imgt.org) correlates with (ilmfi and that ol' (:1 l1
`lysine (lMCT K120) with (ilml7. The presence ofCl 1.”) aspar—
`tare 556 (iMGT 1)]2) and lcucirre 558 (1M(i'11 1.1/1) correlates
`with (.ilrnl.
`In contrast.
`the presence of (£113 glutamate and
`methionine at 356 arid 558 ilMG’l' 1'112, M14), as observed in
`
`Human immunoglohulin allotypes
`
`Heavy gamma] chains: {Elm}, (ilnrl7.i or (:lnrl7,|,2
`
`Kappa light chains: Kml, Krnl.2 or Km}
`
`Figure 1 Correlations between 196 Gim and Km allotypes and amino
`acids.
`
`Table 1 Human immunoglobulin ullotypes
`
`lgA
`
`AZm
`
`1
`to
`
`light chains
`
`K
`
`Km
`
`Lars)—
`
`Isotype/type
`
`IgU |
`
`Allotypes
`
`(ilm
`
`so)
`
`Heavy chains
`1g(12
`1g(13
`
`(i3m
`
`Zligl)
`ZXigS)
`litho)
`Sibl)
`13(b3)
`Mitt-’1)
`iitihS)
`15is)
`1611)
`6(03)
`24te5)
`Hutu)
`27H)
`
`NB: Alphdtgtal notation given within bratkets,
`
`sera of patients with multiple rnycloma. He immunized rabbits
`with these proteins, used cross—absorption experiments to show
`that each expressed unique antigenic determinants (epitopes)
`and coined the term ”individual antigenic specificity" (1A8);
`he distinguished this phenomenon from idiotypy because the
`serologic reagents were generated by xeno—immuni/.ation, not allo—
`immunization.” With the passage of‘time, both phenomena were
`referred to as idiotypy. In the current context, mAb treatment of
`patients may lead to both allo—immunization and/or xeno—immuni—
`zation that results in antisera tirat may recognize isotypic, allotypic
`and idiotypic epitopcs. Whilst it is generally held that the idiotype
`reflects the uniqueness of the combined VH and VL sequences,
`there is evidence that the idiotype can be influenced by the heavy
`Chain constant region with which they are expressetl.2”'“
`
`334
`
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`
`2009; Vol.
`
`1 lssue4
`
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`

`Human immunoglobulin ullolypes
`
`Heat) gaitimaleltaiiis: (.21111301‘(.'11111..)
`
`Kappa light chains: K1111.l\'1111.211rl\'1113
`
`1‘1419101
`
`(311111")
`
`\
`
`\c
`
`T
`T
`\ I
`|\111|
`11 ‘l‘iiii
`kuil‘2\l,rl '1‘11
`Km]
`
`Figure 2. Correlations between lgG (32m and Km ullolypes and amino
`acids
`
`Table 2 Prevalent Gm haplofypes in different populations
`
`ofCHZ ntethionine at residue 282 (IMGT M431), Fig. 2. The
`(3211123 chains are also characteriled by the presence of CH1
`tbreonine 189 (IMGT T92).3'4 The GZm(..) or GZni(n—) chains
`and gamma chains ofother lgG subclasses have CH1 ptolitte 189
`(lMGT 92) and CH2 valine 282 (IMGT V431); however, these
`amino acid residues do not seem immunogenic and cannot be
`defined as isoallotypes as no mAb reagent has been characterized
`lgG3.
`It
`is evident from Table 1
`that serologically defined
`allotypy within the IgGfi subclass is very complex. Assigning
`sequence correlates to 1gG3 proteins expressing a given cltister of
`allotypes had to await dedicated analysis of multiple 1gG3 protein
`sequences. The result of such analysis is summarized in Table 3;
`however, there is another layer of complexity that underlies these
`assignments. Thus, analysis of 19 DNA sequences of the G3mS”
`or G3ni(b*) haplotype yielded 11 different allelic sequences; simi—
`larly analysis of 10 DNA sequences of the G311121" or G3in(g1*)
`liaplotype yielded 4 distinct sequences.35 It is not known whether
`amino acid changes dtie to these polymorphisms are immunogenic
`and could, potentially, be detected by uniqtte serological reagents,
`An additional polymorphism results
`from differing numbers
`7
`of binge region exons. The hinge may be encoded
`by 2~5 exoiis;
`the first encoding a common hinge
`region with 1—4 repeats ofa second exon.3(‘ Thus, the
`hinge region protein sequence can vary from 27 to
`,
`.
`.
`.
`i
`.
`85 amino acid residues, and cait influence structural
`-
`-
`characteristics.
`1gG4. No allotypes have been defined for 1gG4.
`The setologically defined lgG4 polymorphism
`being the isoallotypes ttG4nt(a) and 11G4m(b). The
`nG4m(a) cpitope that C()rrelates with CH2 leucine
`‘
`\
`309 (lMGT L92) is also expressed on 1gb] and IgG3
`~
`,
`‘
`_
`heavy constant regions, whereas the 11(14m(l'))'cpll()pc
`that correlates with CH2 valine 309 (IMC: T V92)
`is also expressed on the lgGZ heavy chain constant
`region.
`Kappa and lambda light chains. The lgGK/lgGJ»
`expression ratio is approximately 60/40. The human
`genome has one kappa constant (IGKC) gene btit variable number
`ol'ilambda constant (IGLC) genes; therefore, we need to consider
`two different forms of polymorphism, eaclt of which may have
`consequences for immtinogenicity oi‘mAbs. There are three kappa
`chain allotypes designated Kml, K1112 and K1113 that define three
`Km alleles: Kml, that correlates with valinelS3 (IMGT V451)
`and leticiite 191 (IMGT L101), Kml,2 that correlates with alanine
`153 (IMGT A431 and leucine 191 and K1113 that correlates with
`alanine 153 and valine 191 (IMGT V101)”
`Serologically defined allotypes of the constant region of lambda
`chains have not been reported; however,
`the number of IGLC
`genes varies between 7 and 11 depending on individual haplo—
`typeslii'jx’qU These isotypes can also be distinguished serologically.
`lit
`the 7—geiie haplotype lGLCl,
`lGLCZ, 1(3LC3 and IGLC7
`are lunctional, lGLC/t and IGLCS are pseudogenes and 1GLC6
`is either lunctional iii a rare haplotype” or, more frequently, a
`psetidogene