The history of isotype selection for therapeutic antibodies clearly demonstrates that solid understanding of the biologic effects mediated by immunoglobulin (Ig) subclasses IgG1, IgG2, IgG3, and IgG4 and their variants is a prerequisite for optimal antibody development. The choices of isotype-constant regions for full-length antibody generation are becoming more complex by the availability of engineered variants and, in some cases, mixed isotypes. This review focuses on important lessons from preclinical studies, and clinical trials, which underscore that careful selection of isotypes is critical given their differential effects on antibody function. The molecular requirements vary for different mechanisms of action of therapeutic antibodies (eg, neutralizing, agonistic, conjugated), and IgG properties beneficial for one mechanism might be detrimental for another. Key isotype selection principles for optimal antibody development are reviewed, and case studies are discussed.
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References
Anderson D, Chambers K, Hanna N, Leonard J, Reff M, Newman R, Baldoni J, Dunleavy D, Reddy M, Sweet R, Truneh A (1997) A primatized MAb to human CD4 causes receptor modulation without marked reduction in CD4+ T cells in chimpanzees: in vitro and in vivo characterization of a MAb (IDEC-CE9.1) to human CD4. Clin Immunol Immunopathol 84:73–84
Beenhouwer DO, Yoo EM, Lai CW, Rocha MA, Morrison SL (2007) Human immunoglobulin G2 (IgG2) and IgG4, but not IgG1 or IgG3, protect mice against Cryptococcus neoformans infection. Infect Immun 75:1424–1435
Blaich G, Janssen B, Roth G, Salfeld J (2007) Overview: differentiating issues in the development of macromolecules compared with small molecules. In: Gad SC (ed) Handbook of pharmaceutical biotechnology. John Wiley & Sons, Hoboken, pp 89–123
Bowles JA, Weiner GJ (2005) CD16 polymorphisms and NK activation induced by monoclonal antibody-coated target cells. J Immunol Methods 304:88–99
Canfield SM, Morrison SL (1991) The binding affinity of human IgG for its high affinity Fc receptor is determined by multiple amino acids in the CH2 domain and is modulated by the hinge region. J Exp Med 173:1483–1491
Carter P, Presta L, Gorman CM, Ridgway JB, Henner D, Wong WL, Rowland AM, Kotts C, Carver ME, Shepard HM (1992) Humanization of an anti-p185HER2 antibody for human cancer therapy. Proc Natl Acad Sci USA 89:4285–4289
Casadevall A, Cleare W, Feldmesser M, Glatman-Freedman A, Goldman DL, Kozel TR, Lendvai N, Mukherjee J, Pirofski LA, Rivera J, Rosas AL, Scharff MD, Valadon P, Westin K, Zhong Z (1998) Characterization of a murine monoclonal antibody to Cryptococcus neoformans polysaccharide that is a candidate for human therapeutic studies. Antimicrob Agents Chemother 42:1437–1446
Chamberlain AK, Desjarlais JR, Karki SB, Lazar GA, Vielmetter J, Yoder SC (2006) inventors; Xencor, Inc, assignee. Fc variants with altered binding to FcRn. US patent application 20,070,135,620
Chatenoud L, Ferran C, Legendre C, Thouard I, Merite S, Reuter A, Gevaert Y, Kreis H, Franchimont P, Bach JF (1990) In vivo cell activation following OKT3 administration. Systemic cytokine release and modulation by corticosteroids. Transplantation 49:697–702
Dall’Acqua WF, Cook KE, Damschroder MM, Woods RM, Wu H (2006) Modulation of the effector functions of a human IgG1 through engineering of its hinge region. J Immunol 177:1129–1138
Dillon TM, Ricci MS, Vezina C, Flynn GC, Liu YD, Rehder DS, Plant M, Henkle B, Li Y, Deechongkit S, Varnum B, Wypych J, Balland A, Bondarenko PV (2008) Structural and functional characterization of disulfide isoforms of the human IgG2 subclass. J Biol Chem 283:16206–16215
Duncan AR, Winter G (1988) The binding site for C1q on IgG. Nature 332:738–740
Gillies SD, Lan Y, Williams S, Carr F, Forman S, Raubitschek A, Lo KM (2005) An anti-CD20-IL-2 immunocytokine is highly efficacious in a SCID mouse model of established human B lymphoma. Blood 105:3972–3978
Greenwood J, Clark M, Waldmann H (1993) Structural motifs involved in human IgG antibody effector functions. Eur J Immunol 23:1098–1104
Guler-Gane G, Holgate R, Jermutus L, Levens J, Lund J, Stewart R, Thom A, Webster C (2008) inventors; MedImmune, assignee. Fc polypeptide variants obtained by ribosome display methodology. International patent application WO/2008/114011
Hale G, Bright S, Chumbley G, Hoang T, Metcalf D, Munro AJ, Waldmann H (1983) Removal of T cells from bone marrow for transplantation: a monoclonal antilymphocyte antibody that fixes human complement. Blood 62:873–882
Hale G, Dyer MJ, Clark MR, Phillips JM, Marcus R, Riechmann L, Winter G, Waldmann H (1988) Remission induction in non-Hodgkin lymphoma with reshaped human monoclonal antibody CAMPATH-1H. Lancet 2:1394–1399
Hepburn TW, Totoritis MC, Davis CB (2003) Antibody-mediated stripping of CD4 from lymphocyte cell surface in patients with rheumatoid arthritis. Rheumatology (Oxford) 42:54–61
Herzog C, Walker C, Müller W, Rieber P, Reiter C, Riethmüller G, Wassmer P, Stockinger H, Madic O, Pichler WJ (1989) Anti-CD4 antibody treatment of patients with rheumatoid arthritis: I. Effect on clinical course and circulating T cells. J Autoimmun 2:627–642
Hinton PR, Tsurushita N, Tso JY, Vasquez M, inventors; PDL BioPharma (2008) Inc, assignee. Alteration of FcRn binding affinities or serum half-lives of antibodies by mutagenesis. US patent 7,361,740
HUMIRA Biologic License Application (BLA 125057). Abbott Laboratories, Abbott Park, http://www.fda.gov/cder/biologics/review/BLA125057_S000_HUMIRA_PHARMR.pdf. Accessed 11 Feb 2009
Idusogie EE, Presta LG, Mulkerrin MG (2001) inventors; Genentech, Inc, assignee. Polypeptide variants. US patent 6194551
International Immunogenetics Information System. Gene table: Human (Homo sapiens) IGHC. http://www.imgt.org/textes/IMGTrepertoire/LocusGenes/tabgenes/human/IGH/IGHC/Hu_IGHC.html. Cited 22 Jan 2009
Isaacs JD, Wing MG, Greenwood JD, Hazleman BL, Hale G, Waldmann H (1996) A therapeutic human IgG4 monoclonal antibody that depletes target cells in humans. Clin Exp Immunol 106:427–433
Jefferis R, Lund J, Goodall M (1995) Recognition sites on human IgG for Fc gamma receptors: the role of glycosylation. Immunol Lett 44:111–117
Jolliffe LK (1993) Humanized antibodies: enhancing therapeutic utility through antibody engineering. Int Rev Immunol 10:241–250
Kon OM, Sihra BS, Compton CH, Leonard TB, Kay AB, Barnes NC (1998) Randomised, dose-ranging, placebo-controlled study of chimeric antibody to CD4 (keliximab) in chronic severe asthma. Lancet 352:1109–1113
Kung P, Goldstein G, Reinherz EL, Schlossman SF (1979) Monoclonal antibodies defining distinctive human T cell surface antigens. Science 206:347–349
Langer F, Ingersoll SB, Amirkhosravi A, Meyer T, Siddiqui FA, Ahmad S, Walker JM, Amaya M, Desai H, Francis JL (2005) The role of CD40 in CD40L- and antibody-mediated platelet activation. Thromb Haemost 93:1137–1146
Lazar GA, Chirino AJ, Wei D, Desjarlais JR, Doberstein SK, Hayes RJ, Karki SB, Vafa O (2008) inventors; Xencor, Inc, assignee. Optimized Fc variants. US patent 7317091
Lendvai N, Qu XW, Hsueh W, Casadevall A (2000) Mechanism for the isotype dependence of antibody-mediated toxicity in Cryptococcus neoformans-infected mice. J Immunol 164:4367–4374
Liu YD, Chen X, Enk JZ, Plant M, Dillon TM, Flynn GC (2008) Human IgG2 antibody disulfide rearrangement in vivo. J Biol Chem 283:29266–29272
Manches O, Lui G, Chaperot L, Gressin R, Molens JP, Jacob MC, Sotto JJ, Leroux D, Bensa JC, Plumas J (2003) In vitro mechanisms of action of rituximab on primary non-Hodgkin lymphomas. Blood 101:949–954
McDonagh CF, Kim KM, Turcott E, Brown LL, Westendorf L, Feist T, Sussman D, Stone I, Anderson M, Miyamoto J, Lyon R, Alley SC, Gerber HP, Carter PJ (2008) Engineered anti-CD70 antibody-drug conjugate with increased therapeutic index. Mol Cancer Ther 7:2913–2923
McLean GR, Torres M, Elguezabal N, Nakouzi A, Casadevall A (2002) Isotype can affect the fine specificity of an antibody for a polysaccharide antigen. J Immunol 169:1379–1386
Mould DR, Davis CB, Minthorn EA, Kwok DC, Elliott MJ, Luggen ME, Totoritis MC (1999) A population pharmacokinetic-pharmacodynamic analysis of single doses of clenoliximab in patients with rheumatoid arthritis. Clin Pharmacol Ther 66:246–257
Natsume A, In M, Takamura H, Nakagawa T, Shimizu Y, Kitajima K, Wakitani M, Ohta S, Satoh M, Shitara K, Niwa R (2008) Engineered antibodies of IgG1/IgG3 mixed isotype with enhanced cytotoxic activities. Cancer Res 68:3863–3872
Newman R, Hariharan K, Reff M, Anderson DR, Braslawsky G, Santoro D, Hanna N, Bugelski PJ, Brigham-Burke M, Crysler C, Gagnon RC, Dal Monte P, Doyle ML, Hensley PC, Reddy MP, Sweet RW, Truneh A (2001) Modification of the Fc region of a primatized IgG antibody to human CD4 retains its ability to modulate CD4 receptors but does not deplete CD4(+) T cells in chimpanzees. Clin Immunol 98:164–174
Nussbaum G, Cleare W, Casadevall A, Scharff MD, Valadon P (1997) Epitope location in the Cryptococcus neoformans capsule is a determinant of antibody efficacy. J Exp Med 185:685–694
Pop LM, Liu X, Ghetie V, Vitetta ES (2005) The generation of immunotoxins using chimeric anti-CD22 antibodies containing mutations which alter their serum half-life. Int Immunopharmacol 5:1279–1290
Prahl JW (1967) The C-terminal sequences of the heavy chains of human immunoglobulin G myeloma proteins of differing isotopes and allotypes. Biochem J 105:1019–1028
Presta L (2008) inventor; Genentech, Inc, assignee. Polypeptide variants with altered effector functions. US patent 7371826
Puan KJ, Jin C, Wang H, Sarikonda G, Raker AM, Lee HK, Samuelson MI, Märker-Hermann E, Pasa-Tolic L, Nieves E, Giner JL, Kuzuyama T, Morita CT (2007) Preferential recognition of a microbial metabolite by human Vgamma2Vdelta2 T cells. Int Immunol 19:657–673
Reddy MP, Kinney CA, Chaikin MA, Payne A, Fishman-Lobell J, Tsui P, Dal Monte PR, Doyle ML, Brigham-Burke MR, Anderson D, Reff M, Newman R, Hanna N, Sweet RW, Truneh A (2000) Elimination of Fc receptor-dependent effector functions of a modified IgG4 monoclonal antibody to human CD4. J Immunol 164:1925–1933
Reff ME, Carner K, Chambers KS, Chinn PC, Leonard JE, Raab R, Newman RA, Hanna N, Anderson DR (1994) Depletion of B cells in vivo by a chimeric mouse human monoclonal antibody to CD20. Blood 83:435–445
Reichert JM (2008) Monoclonal antibodies as innovative therapeutics. Curr Pharm Biotechnol 9:423–430
Salfeld JG (2007) Isotype selection in antibody engineering. Nat Biotechnol 25:1369–1372
Shan D, Ledbetter JA, Press OW (1998) Apoptosis of malignant human B cells by ligation of CD20 with monoclonal antibodies. Blood 91:1644–1652
Sharma A, Davis CB, Tobia LA, Kwok DC, Tucci MG, Gore ER, Herzyk DJ, Hart TK (2000) Comparative pharmacodynamics of keliximab and clenoliximab in transgenic mice bearing human CD4. J Pharmacol Exp Ther 293:33–41
Shields RL, Namenuk AK, Hong K, Meng YG, Rae J, Briggs J, Xie D, Lai J, Stadlen A, Li B, Fox JA, Presta LG (2001) High resolution mapping of the binding site on human IgG1 for Fc gamma RI, Fc gamma RII, Fc gamma RIII, and FcRn and design of IgG1 variants with improved binding to the Fc gamma R. J Biol Chem 276:6591–6604
Stavenhagen J, Vijh S, Rankin C, Gorlatov S, Huang L (2008) inventors; Macrogenics, Inc, assignee. Identification and engineering of antibodies with variant Fc regions and methods of using same. US patent 7,355,008
Suitters AJ, Foulkes R, Opal SM, Palardy JE, Emtage JS, Rolfe M, Stephens S, Morgan A, Holt AR, Chaplin LC, Shaw NE, Nesbitt AM, Bodmer MW (1994) Differential effect of isotype on efficacy of anti-tumor necrosis factor alpha chimeric antibodies in experimental septic shock. J Exp Med 179:849–856
Summary of Tysabri Post-Marketing Commitments. http://www.investorvillage.com/smbd.asp?mb=160&mn=76891&pt=msg&mid=1383292. Cited 22 Jan 2009
Uchida J, Hamaguchi Y, Oliver JA, Ravetch JV, Poe JC, Haas KM, Tedder TF (2004) The innate mononuclear phagocyte network depletes B lymphocytes through Fc receptor-dependent mechanisms during anti-CD20 antibody immunotherapy. J Exp Med 199:1659–1669
Valabrega G, Montemurro F, Aglietta M (2007) Trastuzumab: mechanism of action, resistance and future perspectives in HER2-overexpressing breast cancer. Ann Oncol 18:977–984
van de Winkel J, Vink T, Schuurman J, Parren P, Aalberse R, van der Neut Kolfshoten M (2008) inventors; Genmab A/S, assignee. Stable IgG4 antibodies. International patent application WO/2008/145142
van der Lubbe PA, Reiter C, Breedveld FC, Krüger K, Schattenkirchner M, Sanders ME, Riethmüller G (1993) Chimeric CD4 monoclonal antibody cM-T412 as a therapeutic approach to rheumatoid arthritis. Arthritis Rheum 36:1375–1379
van der Neut Kolfschoten M, Schuurman J, Losen M, Bleeker WK, MartÃnez-MartÃnez P, Vermeulen E, den Bleker TH, Wiegman L, Vink T, Aarden LA, De Baets MH, van de Winkel JG, Aalberse RC, Parren PW (2007) Anti-inflammatory activity of human IgG4 antibodies by dynamic Fab arm exchange. Science 317:1554–1557
Vitetta ES, Stone M, Amlot P, Fay J, May R, Till M, Newman J, Clark P, Collins R, Cunningham D, Ghetie V, Uhr JW, Thorpe PE (1991) Phase I immunotoxin trial in patients with B-cell lymphoma. Cancer Res 51:4052–4058
Wagner K, Schulz P, Scholz A, Wiedenmann B, Menrad A (2008) The targeted immunocytokine L19-IL2 efficiently inhibits the growth of orthotopic pancreatic cancer. Clin Cancer Res 14:4951–4960
Wang SY, Weiner G (2008) Complement and cellular cytotoxicity in antibody therapy of cancer. Expert Opin Biol Ther 8:759–768
Wing MG, Moreau T, Greenwood J, Smith RM, Hale G, Isaacs J, Waldmann H, Lachmann PJ, Compston A (1996) Mechanism of first-dose cytokine-release syndrome by CAMPATH 1-H: involvement of CD16 (FcgammaRIII) and CD11a/CD18 (LFA-1) on NK cells. J Clin Invest 98:2819–2826
Woodle ES, Thistlethwaite JR, Jolliffe LK, Zivin RA, Collins A, Adair JR, Bodmer M, Athwal D, Alegre ML, Bluestone JA (1992) Humanized OKT3 antibodies: successful transfer of immune modulating properties and idiotype expression. J Immunol 148:2756–2763
Woodle ES, Xu D, Zivin RA, Auger J, Charette J, O’Laughlin R, Peace D, Jollife LK, Haverty T, Bluestone JA, Thistlethwaite JR Jr (1999) Phase I trial of a humanized, Fc receptor nonbinding OKT3 antibody, huOKT3gamma1(Ala-Ala) in the treatment of acute renal allograft rejection. Transplantation 68:608–616
Wu AM, Senter PD (2005) Arming antibodies: prospects and challenges for immunoconjugates. Nat Biotechnol 23:1137–1146
Wypych J, Li M, Guo A, Zhang Z, Martinez T, Allen MJ, Fodor S, Kelner DN, Flynn GC, Liu YD, Bondarenko PV, Ricci MS, Dillon TM, Balland A (2008) Human IgG2 antibodies display disulfide-mediated structural isoforms. J Biol Chem 283:16194–16205
Xu D, Alegre ML, Varga SS, Rothermel AL, Collins AM, Pulito VL, Hanna LS, Dolan KP, Parren PW, Bluestone JA, Jolliffe LK, Zivin RA (2000) In vitro characterization of five humanized OKT3 effector function variant antibodies. Cell Immunol 200:16–26
Yakes FM, Chinratanalab W, Ritter CA, King W, Seelig S, Arteaga CL (2002) Herceptin-induced inhibition of phosphatidylinositol-3 kinase and Akt Is required for antibody-mediated effects on p27, cyclin D1, and antitumor action. Cancer Res 62:4132–4141
Yednock T (2007) inventor; Elan Pharmaceuticals, Inc, assignee. Methods of treating inflammatory and autoimmune diseases with natalizumab. International patent application WO/2007/103112
Yoo EM, Wims LA, Chan LA, Morrison SL (2003) Human IgG2 can form covalent dimers. J Immunol 170:3134–3138
Zebedee SL, Koduri RK, Mukherjee J, Mukherjee S, Lee S, Sauer DF, Scharff MD, Casadevall A (1994) Mouse-human immunoglobulin G1 chimeric antibodies with activities against Cryptococcus neoformans. Antimicrob Agents Chemother 38:1507–1514
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Editorial support was provided by Robin L. Stromberg, PhD, of Arbor Communications, Inc. (Ann Arbor, MI, USA) and funded by Abbott Laboratories.
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Kaymakcalan, Z., Ibraghimov, A., Goodearl, A.G., Salfeld, J.G. (2010). Aspects of Isotype Selection. In: Kontermann, R., Dübel, S. (eds) Antibody Engineering. Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-01144-3_19
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