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Bispecific Antibodies: Developments and Current Perspectives

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Bispecific Antibodies

Abstract

All naturally occurring antibodies are multifunctional molecules, combining antigen-binding activity and Fc-mediated effector functions within a single molecule (Schroeder and Cavacini 2010). Antigen binding can lead to direct neutralization of the antigen, i.e., being antagonistic, but can also have agonistic activities, e.g., through activation of receptors (Fig.1.1). The Fc region is capable of mediating further effector functions, which include antibody-dependent cellular cytotoxicity (ADCC), complement-dependent cytolysis (CDC) and opsonization, antibody-dependent cellular phagocytosis (ADCP), degranulation, antigen presentation, plasma recycling processes, and regulation of cell activation and proliferation (Fig.1.1). Consequently, natural antibodies are well suited for a variety of therapeutic applications utilizing one or more of these effector functions. These effector functions can be further improved by protein and/or glyco-engineering (Carter 2006; Kubota et al. 2009). However, natural antibodies also face certain limitations. For example, antibodies are not capable of recruiting T cells because these cells do not express Fc receptors. Also, monoclonal antibodies are monospecific, thus, cannot address different target. Hence, strategies have been developed to extend the effector functions to novel activities normally not associated with antibodies, for example, by direct conjugation or fusion of therapeutic compounds or by indirect means using bispecific antibodies.

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References

  • Alt M, Müller R, Kontermann RE (1999) Novel tetravalent and bispecific IgG-like antibody molecules combining single-chain diabodies with the immunoglobulin g1 Fc or CH3 region. FEBS Lett 454:90–94

    Article  PubMed  CAS  Google Scholar 

  • Amoroso RA, Alpaugh RK, Barth MW, McCall AM, Weiner LM (1999) Production and characterization of mice transgenic for the A and B isoforms of human FcgRIII. Cancer Immunol Immunother 48:443–455

    Article  PubMed  CAS  Google Scholar 

  • Asano R, Watanabe Y, Kawaguchi H, Fukazawa H, Nakanishi T, Umetsu M, Hayasi H, Katayose Y, Unno M, Kudo T, Kumagai I (2007) Highly effective recombinant format of a humanized IgG-like bispecific antibody for cancer immunotherapy with retargeting of lymphocytes to tumor cells. J Biol Chem 282:27659–27665

    Article  PubMed  CAS  Google Scholar 

  • Atwell S, Ridgway JBB, Wells JA, Carter P (1997) Stable heterodimers from remodelling the domain interface of a homodimer using a phage display library. J Mol Biol 270:26–35

    Article  PubMed  CAS  Google Scholar 

  • Biburger M, Weth R, Wels WS (2005) A novel bispecific tetravalent antibody fusion protein to target costimulatory activity for T-cell activation to tumor cells overexpressing ErbB2/HER2. J Mol Biol 346:1299–1311

    Article  PubMed  CAS  Google Scholar 

  • Bostrom J, Yu SF, Kan D, Appleton BA, Lee CV, Billeci K, man W, Peale F, Ross S, Wiesmann C, Fuh G (2009) Variants of the antibody herceptin that interacts with HER2 and VEGF at the antigen binding site. Science 323:1610–1614

    Article  PubMed  CAS  Google Scholar 

  • Brandão JG, Scheper RJ, Lougheed SM, Curiel DT, Tillman BW, Gerritsen WR, van den Eertwegh AJ, Pinedo HM, Haisma HJ, de Gruijl TD (2003) CD40-targeted adenoviral gene transfer to dendritic cells through the use of a novel bispecific single-chain Fv antibody enhances cytotoxic T cell activation. Vaccine 21:2268–2272

    Article  PubMed  CAS  Google Scholar 

  • Brekke OH, Sandlie I (2003) Therapeutic antibodies for human diseases at the dawn of the twenty-first century. Nat Rev Drug Discov 22:52–62

    Article  CAS  Google Scholar 

  • Brennan M, Davison PF, Paulus H (1985) Preparation of bispecific antibodies by chemical recombination of monoclonal immunoglobulin G1 fragments. Science 229:81–83

    Article  PubMed  CAS  Google Scholar 

  • Brischwein K, Parr L, Pflanz S, Volkland J, Lumsden J, Klinger M, Locher M, Hammond SA, Kiener P, Kufer P, Schlereth B, Baeuerle PA (2007) Strictly target cell-dependent activation of T cells by bispecific single-chain antibody constructs of the BiTE class. J Immunother 30:798–807

    Article  PubMed  CAS  Google Scholar 

  • Bruenke J, Fischer B, Barbin K, Schreiter K, Wachter Y, Mahr K, Titgemeyer F, Niederweis M, Peipp M, Zunino S, Repp R, Valerius T, Fey GH (2004) A recombinant bispecific single-chain Fv antibody against HLA class II and FcγRIII (CD16) triggers effective lysis of lymphoma cells. Br J Haematol 125:167–179

    Article  PubMed  CAS  Google Scholar 

  • Bruenke J, Barbin K, Kunert S, Lang P, Pfeiffer M, Stieglmaier K, Niethammer D, Stockmeyer B, Peipp M, Repp R, Valerius T, Fey GH (2005) Effective lysis of lymphoma cells with a stabilised bispecific single-chain Fv antibody against CD19 and FcgammaRIII (CD16). Br J Haematol 130:218–228

    Article  PubMed  CAS  Google Scholar 

  • Brüsselbach S, Korn T, Völkel T, Müller R, Kontermann RE (1999) Enzyme recruitment and tumor cell killing in vitro by a secreted bispecific single-chain diabody. Tumor Target 4:115–123

    Google Scholar 

  • Bubenik J (2003) Tumor MHC class I downregulation and immunotherapy. Oncol Rep 10:2205–2208

    Google Scholar 

  • Cao Y, Lam L (2003) Bispecific antibody conjugates in therapeutics. Adv Drug Deliv Rev 55:171–197

    Article  PubMed  CAS  Google Scholar 

  • Cao Y, Suresh MR (1998) Bispecific antibodies as novel bioconjugates. Bioconjug Chem 9:635–644

    Article  PubMed  Google Scholar 

  • Carter P (2001) Bispecific human IgG by design. J Immunol Meth 248:7–15

    Article  CAS  Google Scholar 

  • Carter PJ (2006) Potent antibody therapeutics by design. Nat Rev Immunol 6:343–357

    Article  PubMed  CAS  Google Scholar 

  • Chan AC, Carter PJ (2010) Therapeutic antibodies for autoimmunity and inflammation. Nat Rev Immunol 10:301–316

    Article  PubMed  CAS  Google Scholar 

  • Chang CH, Sharkey RM, Rossi EA, Karacay H, McBride W, Hansen HJ, Chatal JF, Barbet J, Goldenberg DM (2002) Molecular advances in pretargeting radioimmunotherapy with bispecific antibodies. Mol Cancer 1:553–563

    CAS  Google Scholar 

  • Cochlovius B, Kipriyanov SM, Stassar MJJG, Schuhmacher J, Benner A, Moldenhauer G, Little M (2000) Cure of Burkitt’s lymphoma in severe combined immunodeficiency mice by T cells, tetravalent CD3 x CD19 tandem diabody, and CD28 costimulation. Cancer Res 60:4336–4341

    PubMed  CAS  Google Scholar 

  • Coloma MJ, Morrison SL (1997) Design and production of novel tetravalent bispecific antibodies. Nat Biotechnol 15:159–163

    Article  PubMed  CAS  Google Scholar 

  • Connelly RJ, Hayden MS, Scholler JK, Tsu TT, Dupont B, Ledbetter JA, Kanner SB (1998) Mitogenic properties of a bispecific single-chain Fv-Ig fusion generated from CD2-specific mAb to distinct epitopes. Int Immunol 10:1863–1872

    Article  PubMed  CAS  Google Scholar 

  • Corvalan JR, Smith W (1987) Construction and characterisation of a hybrid-hybrid monoclonal antibody recognising both carcinoembryonic antigen (CEA) and vinca alkaloids. Cancer Immunol Immunother 24:127–132

    PubMed  CAS  Google Scholar 

  • Curnow RT (1997) Clinical experience with CD64-directed immunotherapy. An overview. Cancer Immunol Immunother 45:210–215

    Article  PubMed  CAS  Google Scholar 

  • de Gast GC, van de Winkel JGJ, Bast BEJEG (1997) Clinical perspectives of bispecific antibodies in cancer. Cancer Immunol Immunother 45:121–123

    Article  PubMed  Google Scholar 

  • de Kruif J, Logtenberg T (1996) Leucine zipper dimerized bivalent and bispecific scFv antibodies from a semisynthetic antibody phage display library. J Biol Chem 271:7630–7634

    Article  PubMed  Google Scholar 

  • de Lau WB, Keije K, Neefjes JJ, Oosterwegel M, Rozemüller E, Blast BJ (1991) Absence of preferential homologous H/L chain association in hybrid hybridomas. J Immunol 146:906–914

    PubMed  Google Scholar 

  • Deo YM, Sundarapandiyan K, Keler T, Wallace PK, Graziano RF (1998) Bispecific molecules directed to the Fc receptor for IgA (FcαRI, CD89) and tumor antigens efficiently promote cell-mediated cytotoxicity of tumor targets in whole blood. J Immunol 160:1677–1686

    PubMed  CAS  Google Scholar 

  • Dimasi N, Gao C, Fleming R, Woods RM, Yao X, Shirinian L, Kiener PA, Wu H (2009) The design and characterization of oligospecific antibodies for simultaneous targeting of multiple disease mediators. J Mol Biol 393:672–692

    Article  PubMed  CAS  Google Scholar 

  • Dreier T, Baeuerle PA, Fichtner I, Grün M, Schlereth B, Lorenczewski G, Kufer P, Lutterbüse R, Riethmüller G, Gjorstrup P, Bargou RC (2003) T cell costimulus-independent and very efficacious inhibition of tumor growth in mice bearing subcutaneous or leukemic human B cell lymphoma xenografts by a CD19-/CD3-bispecific single-chain antibody construct. J Immunol 170:4397–4402

    PubMed  CAS  Google Scholar 

  • Fanger MW, Segal DM, Romet-Lemonne J-L (1991) Bispecific antibodies and targeted cellular cytotoxicity. Immunol Today 12:51–54

    Article  PubMed  CAS  Google Scholar 

  • Fanger MW, Morganelli PM, Guyre PM (1992) Bispecific antibodies. Crit Rev Immunol 12:101–124

    PubMed  CAS  Google Scholar 

  • FitzGerald K, Holliger P, Winter G (1997) Improved tumor targeting by disulphide stabilized diabodies expressed in Pichia pastoris. Protein Eng 10:1221–1225

    Article  PubMed  CAS  Google Scholar 

  • Fury MG, Lipton A, Smith KM, Winston CB, Pfister DG (2008) A phase-I trial of epidermal growth factor receptor directed bispecific antibody MDX-447 without and with recombinant human granulocyte-colony stimulating factor in patients with advanced solid tumors. Cancer Immunol Immunother 57:155–163

    Article  PubMed  CAS  Google Scholar 

  • Gautherot E, Bouhou J, Le Doussal JM, Manetti C, Martin M, Rouvier E, Barbet J (1997) Therapy of colon carcinoma xenografts with bispecific antibody-targeted, iodine-141-labeled bivalent hapten. Cancer 80(Suppl):2618–2623

    Article  PubMed  CAS  Google Scholar 

  • Gerstmayer B, Hoffmann M, Altenschmidt U, Wels W (1997) Costimulation of T-cell proliferation by a chimeric B7-antibody fusion protein. Cancer Immunol Immunother 45:156–158

    Article  PubMed  CAS  Google Scholar 

  • Getts DR, Getts MT, McCarthy DP, Chastain EML, Miller SD (2010) Have we overestimated the benefit of human(ized) antibodies? mAbs 2:682–694

    Article  PubMed  Google Scholar 

  • Glennie MJ, McBride HM, Worth AT, Stevenson GT (1987) Preparation and performance of bispecific F(ab’γ)2 antibody containing thioether-linked Fab’γ fragments. J Immunol 139:2367–2375

    PubMed  CAS  Google Scholar 

  • Goldenberg DM (2003) Advancing role of radiolabeled antibodies in the therapy of cancer. Cancer Immunol Immunother 52:281–296

    PubMed  CAS  Google Scholar 

  • Goldenberg DM, Sharkey RM (2010) Radioactive antibodies: a historical review of selective targeting and treatment of cancer. Hosp Pract 38:82–93

    Article  Google Scholar 

  • Goldenberg DM, Chatal JF, Barbet J, Boerman O, Sharkey RM (2007) Cancer imaging and therapy with bispecific antibody pretargeting. Update Cancer Ther 2:19–31

    Article  PubMed  Google Scholar 

  • Graziano RF, Guptill P (2004) Chemical production of bispecific antibodies. Meth Mol Biol 283:71–85

    CAS  Google Scholar 

  • Grosse-Hovest L, Müller S, Minoia R, Wolf E, Zakhartchenko V, Wenigerkind H, Lassnig C, Besenfelder U, Müller M, Lytton SD, Jung G, Brem G (2004) Cloned transgenic farm animals produce a bispecific antibody for T cell-mediated tumor cell killing. Proc Natl Acad Sci USA 101:6858–6863

    Article  PubMed  CAS  Google Scholar 

  • Gruaz-Guyon A, Raguin O, Barbet J (2005) Recent advances in pretargeted radioimmunotherapy. Curr Med Chem 12:319–338

    Article  PubMed  CAS  Google Scholar 

  • Gunasekaran K, Pentony M, Shen M, Garrett L, Forte C, Woodward A, Ng SB, Born T, Retter M, Mnachulenko K, Sweet H, Foltz IN, Wittekind M, Yan W (2010) Enhancing antibody Fc heterodimer formation through electrostatic steering effects: applications to bispecific molecules and monovalent IgG. J Biol Chem 18:19637–19646

    Article  CAS  Google Scholar 

  • Hartmann F, Renner C, Jung W, Deisting C, Juwana M, Eichentopf B, Kloft M, Pfreundschuh M (1997) Treatment of refractory Hodgkin’s disease with an anti-CD16/CD30 bispecific antibody. Blood 89:2042–2047

    PubMed  CAS  Google Scholar 

  • Hartmann F, Renner C, Jung W, da Costa L, Tembrink S, Held G, Sek A, König J, Bauer S, Kloft M, Pfreundschuh M (2001) Anti-CD16/CD30 bispecific antibody treatment for Hodgkin’s disease: role of infusion schedule and costimulation with cytokines. Clin Cancer Res 7:1873–1881

    PubMed  CAS  Google Scholar 

  • Hayden MS, Linsley PS, Gayle MA, Bajorath J, Brady WA, Norris NA, Fell HP, Ledbetter JA, Gilliland LK (1994) Single-chain mono- and bispecific antibody derivatives with novel biological properties and antitumor activity from a COS cell transient expression system. Ther Immunol 1:3–15

    PubMed  CAS  Google Scholar 

  • Hoffmann P, Hofmeister R, Brischwein K, Brandl C, Crommer S, Bargou R, Itin C, Prang N, Baeuerle PA (2005) Serial killing of tumor cells by cytotoxic T cells redirected with a CD19-/CD3-bispecific single-chain antibody construct. Int J Cancer 115:98–104

    Article  PubMed  CAS  Google Scholar 

  • Holliger P, Prospero T, Winter G (1993) “Diabodies”: small bivalent and bispecific antibody fragments. Proc Natl Acad Sci USA 90:6444–6448

    Article  PubMed  CAS  Google Scholar 

  • Holliger P, Manzke O, Span M, Hawkins R, Fleischmann B, Qianghua L, Wolf J, Diehl V, Cochet O, Winter G, Bohlen H (1999) Carcinoembryonic antigen (CEA)-specific T-cell activation in colon carcinoma induced by anti-CD3xanti-CEA bispecific diabodies and B7xanti-CEA bispecific fusion proteins. Cancer Res 59:2909–2926

    Google Scholar 

  • Hombach A, Jung W, Pohl C, Renner C, Sahin U, Schmits R, Wolf J, Kapp U, Diehl V, Pfreundschuh M (1993) A CD16/CD30 bispecific monoclonal antibody induces lysis of Hodgkin’s cells by unstimulated natural killer cells in vitro and in vivo. Int J Cancer 55:830–836

    Article  PubMed  CAS  Google Scholar 

  • Jackman J, Chen Y, Hhuang A, Moffat B, Scheer JM, Leong SR, Lee WP, Zhang J, Sharma N, Lu Y, Iyer S, Shields RL, Chiang N, Bauer MC, Wadley D, Roose-Girma M, Vandlen R, Yansura DG, Wu Y, Wu LC (2010) Development of two-part strategy to identify a therapeutic human bispecific antibody that inhibits IgE receptor signaling. J Biol Chem 285:20850–20859

    Article  PubMed  CAS  Google Scholar 

  • James ND, Atherton PJ, Jones J, Howie AJ, Tchekmedyian S, Curnow RT (2001) A phase II study of the bispecific antibody MDX-H210 (anti-HER2xCD64) with GM-CSF in HER2+ advanced prostate cancer. B J Cancer 85:152–156

    Article  CAS  Google Scholar 

  • Jimenez X, Lu D, Brennan L, Persaud K, Liu M, Miao H, Witte L, Zhu Z (2005) A recombinant, fully human, bispecific antibody neutralizes the biological activities mediated by both vascular endothelial growth factor receptors 2 and 3. Mol Cancer Ther 4:427–434

    PubMed  CAS  Google Scholar 

  • Johnson S, Burke S, Huang L, Gorlatov S, Lim H, Wangm W, Zhangm W, Tuaillon N, Raineym J, Baratm B, Yangm Y, Jinm L, Ciccaronem V, Moorem PA, Koenigm S, Bonvini E (2010) Effector cell recruitment with novel Fv-based dual-affinity re-targeting protein leads to potent tumor cytolysis and in vivo B-cell depletion. J Mol Biol 399:436–449

    Article  PubMed  CAS  Google Scholar 

  • Kellner C, Bruenke J, Stieglmaier J, Schwemmlein M, Schwenkert M, Singer H, Mentz K, Peipp M, Lang P, Oduncu F, Stockmeyer B, Fey GH (2008) A novel CD19-directed recombinant bispecific antibody derivative with enhanced immune effector functions for human leukemic cells. J Immunother 31:871–884

    Article  PubMed  CAS  Google Scholar 

  • Kipriyanov SM, Moldenhauer G, Schuhmacher J, Cochlovius B, Von der Lieth CW, Matys ER, Little M (1999) Bispecific tandem diabody for tumor therapy with improved antigen binding and pharmacokinetics. J Mol Biol 293:41–56

    Article  PubMed  CAS  Google Scholar 

  • Kipriyanov SM, Cochlovius B, Schäfer HJ, Moldenhauer G, Bähre A, Le Gall F, Knackmuss S, Little M (2002) Synergistic antitumor effect of bispecific CD19xCD3 and CD19xCD16 diabodies in a preclinical model of non-Hodgkin’s lymphoma. J Immunol 169:137–144

    PubMed  CAS  Google Scholar 

  • Köhler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:495–497

    Article  PubMed  Google Scholar 

  • Kontermann RE (2005) Recombinant bispecific antibodies for cancer therapy. Acta Pharmacol Sin 26:1–9

    Article  PubMed  CAS  Google Scholar 

  • Kontermann RE (2009) Strategies to extend plasma half-lives of recombinant antibodies. BioDrugs 23:93–109

    Article  PubMed  CAS  Google Scholar 

  • Kontermann RE (2010) Alternative antibody formats. Curr Opin Mol Ther 12:176–183

    PubMed  CAS  Google Scholar 

  • Kontermann RE, Scheurich P, Pfizenmaier K (2009) Antagonists of TNF action: clinical experience and new developments. Expert Opin Drug Discov 4:279–292

    Article  CAS  Google Scholar 

  • Koolwijk P, Spierenbur GGT, Frasa H, Boot JH, van de Winkel JH, Bat BJ (1989) Interaction between hybrid mouse monoclonal antibodies and the human high-affinity IgG FcR, huFcgRI, on U937. Involvement of only one of the mIgG heavy chains in receptor binding. J Immunol 143:1656–1662

    PubMed  CAS  Google Scholar 

  • Koolwijk P, Boot JH, Griep R, Bast BJ (1991) Binding of human complement subcomponent C1q to hybrid mouse monoclonal antibodies. Mol Immunol 28:567–576

    Article  PubMed  CAS  Google Scholar 

  • Kostelny SA, Cole MS, Tso JY (1992) Formation of a bispecific antibody by the use of leucine zippers. J Immunol 148:1547–1553

    PubMed  CAS  Google Scholar 

  • Kraeber-Bodéré F, Rousseau C, Bodet-Milin C, Ferrer L, Faivre-Chauvet A, Campion L, Vuillez JP, Devillers A, Chang CH, Goldenberg DM, Chatal JF, Barbet J (2006) Targeting, toxicity and efficacy of 2-step, pretargeted radioimmunotherapy using a chimaeric bispecific antibody and 131I-labelled bivalent hapten in a phase I optimization clinical trial. J Nucl Med 47:247–255

    PubMed  Google Scholar 

  • Kranenborg MH, Boeman Oc, Oosterwijk-Wakka JC, de Weijert MC, Corstens FH, Oosterwijk E (1998) Two-step radio-immunotargeting of renal-cell carcinoma xenografts in nude mice with anti-renal-cell-carcinoma X anti-DTPA bispecific monoclonal antibodies. Int J Cancer 75:74–80

    Article  PubMed  CAS  Google Scholar 

  • Kriangkum J, Xu B, Nagata LP, Fulton RE, Suresh MR (2001) Bispecific and bifunctional single chain recombinant antibodies. Biomol Eng 18:31–40

    Article  PubMed  CAS  Google Scholar 

  • Kubota T, Niwa R, Satoh M, Akinaga S, Shitara K, Hanai N (2009) Engineered therapeutic antibodies with improved effector functions. Cancer Sci 1000:1566–1572

    Article  CAS  Google Scholar 

  • Kufer P, Mack M, Gruber R, Lutterbuse R, Zettl F, Riethmuller G (1997) Construction and biological activity of a recombinant bispecific single-chain antibody designed for therapy of minimal residual colorectal cancer. Cancer Immunol Immunother 45:193–197

    Article  PubMed  CAS  Google Scholar 

  • Le Doussal JM, Gruaz-Guyon A, Martin M, Gautherot E, Delaage M, Barbet J (1990) Targeting of indium 111-labeled bivalent hapten to human melanoma mediated by bispecific monoclonal antibody conjugates: imaging of tumors hosted in nude mice. Cancer Res 50:3445–3452

    PubMed  Google Scholar 

  • Lewis LD, Cole BF, Wallace PK, Fisher JL, Waugh M, Guyre PM, Fanger MW, Curnow RT, Kaufman PA, Ernstoff MS (2001) Pharmacokinetic-pharmacodynamic relationship of the bispecific antibody MDX-H210 when administered in combination with interferon gamma: a multiple-dose phase-I study in patients with advanced cancer which overexpresses HER-2/neu. J Immunol Meth 248:149–165

    Article  CAS  Google Scholar 

  • Lindhofer H, Mocika R, Steipe B, Thierfelder S (1995) Preferential species-restricted heavy/light chain pairing in rat/mouse quadromas. Implications for a single-step purification of bispecific antibodies. J Immunol 155:219–225

    PubMed  CAS  Google Scholar 

  • Link BK, Weiner GJ (1993) Production and characterization of a bispecific IgG capable of inducing T-cell-mediated lysis of malignant B cells. Blood 81:3343–3349

    PubMed  CAS  Google Scholar 

  • Lobo ED, Hansen RJ, Balthasar JP (2004) Antibody pharmacokinetics and pharmacodynamics. J Pharm Sci 93:2645–2668

    Article  PubMed  CAS  Google Scholar 

  • Löffler A, Kufer P, Lutterbüse R, Zettl F, Daniel PT, Schwenkenbecher JM, Riethmüller G, Dörken B, Bargou RC (2000) A recombinant bispecific single-chain antibody, CD19 x CD3, induces rapid and high lymphoma-directed cytotoxicity by unstimulated T lymphoctes. Blood 95:2089–2103

    Google Scholar 

  • Lu D, Kotanides H, Jimenez X, Zhou Q, Persaud K, Bohlen P, Witte L, Zhu Z (1999) Acquired antagonistic activity of a bispecific diabody directed against two different epitopes on vascular endothelial growth factor receptor 2. J Immunol Meth 230:159–171

    Article  CAS  Google Scholar 

  • Lu D, Jimenez X, Zhang H, Wu Y, Bohlen P, Witte L, Zhu Z (2001) Complete inhibition of vascular endothelial growth factor (VEGF) activities with a bifunctional diabody directed against both VEGF kinase receptors, fms-like tyrosine kinase receptor and kinase insert domain-containing receptor. Cancer Res 61:7002–7008

    PubMed  CAS  Google Scholar 

  • Lu D, Jimenez X, Zhang H, Bohlen P, Witte L, Zhu Z (2002) Fab-scFv fusion protein: an efficient approach to production of bispecific antibody fragments. J Immunol Meth 267:213–226

    Article  CAS  Google Scholar 

  • Lu D, Jimenez X, Zhang H, Atkins A, Brennan L, Balderes P, Bohlen P, Witte L, Zhu Z (2003) Di-diabody: a novel tetravalent bispecific antibody molecule by design. J Immunol Meth 279:219–232

    Article  CAS  Google Scholar 

  • Lu D, Zhang H, Ludwig D, Persaud A, Jimenez X, Burtrum D, Balderes P, Liu M, Bohlen P, Witte L, Zhu Z (2004) Simultaneous blockade of both the epidermal growth factor receptor and the insulin-like growth factor receptor signaling pathways in cancer cells with a fully human recombinant bispecific antibody. J Biol Chem 279:2856–2865

    Article  PubMed  CAS  Google Scholar 

  • Lu D, Zhang H, Koo H, Tonra J, Balderes P, Prewett M, Corcoran E, Mangalampalli V, Bassi R, Anselma D, Patel D, Kang X, Ludwig DL, Hicklin DJ, Bohlen P, Witte L, Zhu Z (2005) A fully human recombinant IgG-like bispecific antibody to both the epidermal growth factor receptor and the insulin-like growth factor receptor for enhanced antitumor activity. J Biol Chem 280:19665–19672

    Article  PubMed  CAS  Google Scholar 

  • Lum LG, Davol PA (2005) Retargeting T cells and immune effector cells with bispecific antibodies. Cancer Chemother Biol Response Modif 22:273–291

    Article  PubMed  CAS  Google Scholar 

  • Madrenas J, Chau LA, Teft WA, Wu PW, Jussif J, Kasaian M, Carreno BM, Ling V (2004) Conversion of CTLA-4 from inhibitor to activator of T cells with a bispecific tandem single-chain Fv ligand. J Immunol 172:5948–5956

    PubMed  CAS  Google Scholar 

  • Mahmood I, Green MD (2005) Pharmacokinetic and pharmacodynamic considerations in the development of therapeutic proteins. Clin Pharmacokinet 44:331–347

    Article  PubMed  CAS  Google Scholar 

  • Marvin JS, Zhu Z (2005) Recombinant approaches to IgG-like bispecific antibodies. Acta Pharmacol Sin 26:649–658

    Article  PubMed  CAS  Google Scholar 

  • Marvin JS, Zhu Z (2006) Bispecific antibodies for dual-modality cancer therapy: killing two signaling cascades with one stone. Curr Opin Drug Discov Develop 9:184–193

    CAS  Google Scholar 

  • McCall AM, Adams GP, Amoroso AR, Nielsen UB, Zhang L, Horak E, Simmons H, Schier R, Marks JD, Weiner LM (1999) Isolation and characterization of an anti-CD16 single-chain Fv fragment and construction of an anti-HER2/neu/anti-CD16 bispecific scFv that triggers CD16-dependent tumor cytolysis. Mol Immunol 36:433–445

    Article  PubMed  CAS  Google Scholar 

  • McCall AM, Shahied L, Amoroso AR, Horak EM, Simmons HH, Nielson U, Adams GP, Schier R, Marks JD, Weiner LM (2001) Increasing the affinity for tumor antigen enhances bispecific antibody cytotoxicity. J Immunol 166:6112–6117

    PubMed  CAS  Google Scholar 

  • Merchant AM, Zhu Z, Yuan JQ, Goddard A, Adams CW, Presta LG, Carter P (1998) An efficient route to human bispecific IgG. Nat Biotechn 16:677–681

    Article  CAS  Google Scholar 

  • Michaelson JS, Demarest SJ, Miller B, Amatucci A, William S, Wu X, Huang F, Pfan S, Gao S, Doern A et al (2009) Anti-tumor activity of stability-engineered IgG-like bispecific antibodies targeting TRAIL-R2 and LTβR. mAbs 1:128–141

    Article  PubMed  Google Scholar 

  • Milstein C, Cuello AC (1983) Hybrid hybridomas and their use in immunohistochemistry. Nature 305:537–540

    Article  PubMed  CAS  Google Scholar 

  • Morandeau L, Benoist E, Loussouarn A, Quadi A, Lesaec P, Mougin M, Faivre-Chauvet A, Le Boterff J, Chatal JF, Barbet J, Gestin JF (2005) Synthesis of new bivalent peptides for applications in the affinity enhancement system. Bioconjugat Chem 16:184–193

    Article  CAS  Google Scholar 

  • Müller D, Kontermann RE (2007) Recombinant bispecific antibodies for cellular cancer immunotherapy. Curr Opin Mol Ther 9:319–326

    PubMed  Google Scholar 

  • Müller D, Kontermann RE (2010) Bispecific antibodies for cancer immunotherapy: current perspectives. BioDrugs 24:89–98

    Article  PubMed  Google Scholar 

  • Müller KM, Arndt KM, Plückthun A (1998a) A dimeric bispecific miniantibody combines two specificities with avidity. FEBS Lett 432:45–49

    Article  PubMed  Google Scholar 

  • Müller KM, Arndt KM, Strittmatter W, Plückthun A (1998b) The first constant domain (CH1 and CL) of an antibody used as heterodimerisation domain for bispecific miniantibodies. FEBS Lett 422:259–264

    Article  PubMed  Google Scholar 

  • Müller D, Karle A, Meißburger B, Höfig I, Stork R, Kontermann RE (2007) Improved pharmacokinetics of recombinant bispecific antibody molecules by fusion to human serum albumin. J Biol Chem 282:12650–12660

    Article  PubMed  CAS  Google Scholar 

  • Nisonoff A, Rivers MM (1961) Recombination of a mixture of univalent antibody fragments of different specificity. Arch Biochem Biophys 93:460–462

    Article  PubMed  CAS  Google Scholar 

  • Orcutt KD, Ackerman ME, Cieslewicz M, Quiroz E, Slusarczyk AL, Frangioni JV, Wittrup KD (2010) A modular IgG-scFv bispecific antibody topology. Protein Eng Des Sel 23:221–228

    Article  PubMed  CAS  Google Scholar 

  • Pack P, Plückthun A (1992) Miniantibodies: use of amphipathic helices to produce functional, flexibly linked dimeric Fv fragments with high avidity in Escherichia coli. Biochemistry 31:1579–1584

    Article  PubMed  CAS  Google Scholar 

  • Paulus H (1985) Preparation and biomedical applications of bispecific antibodies. Behring Inst Mitt 78:118–132

    PubMed  CAS  Google Scholar 

  • Peipp M, Valerius T (2002) Bispecific antibodies targeting cancer cells. Biochem Soc Trans 30:507–511

    Article  PubMed  CAS  Google Scholar 

  • Plückthun A, Pack P (1997) New protein engineering approaches to multivalent and bispecific antibody fragments. Immunotechnol 3:83–105

    Article  Google Scholar 

  • Pullarkat V, Deo Y, Link J, Spears L, Marty V, Curnow R, Groshen S, Gee C, Weber JS (1999) A phase I study of a HER2/neu bispecific antibody with granulocyte-colony-stimulating factor in patients with metastatic breast cancer that overexpresses HER2/neu. Cancer Immunol Immunother 48:9–21

    Article  PubMed  CAS  Google Scholar 

  • Reilly RM (2006) Radioimmunotherapy of solid tumors: the promise of pretargeting strategies using bispecific antibodies and radiolabelled haptens. J Nuc Med 47:196–199

    CAS  Google Scholar 

  • Ren-Heidenreich L, Davol PA, Kouttab NM, Elfenbein GJ, Lum LG (2004) Redirected T-cell cytotoxicity to epithelial cell adhesion molecule-overexpressing adenocarcinomas by a novel recombinant antibody, E3Bi, in vitro and in an animal model. Cancer 100:1095–1103

    Article  PubMed  CAS  Google Scholar 

  • Repp R, van Ojik HH, Valerius T, Groenewegen G, Wieland G, Oetzel C, Stockmeyer B, Becker W, Eisenhut M, Steininger H, Deo YM, Blijham GH, Kalden JR, van de Winkel JGJ, Gramatzki M (2003) Phase I clinical trial of the bispecific antibody MDX-H210 (anti-FcγRIxanti-HER-2/neu) in combination with Filgrastim (G-CSF) for treatment of advanced breast cancer. B J Cancer 89:2234–2243

    Article  CAS  Google Scholar 

  • Reusch U, Le Gall F, Hensel M, Moldenhauer G, Ho AD, Little M, Kipriyanov SM (2004) Effect of tetravalent bispecific CD19xCD3 recombinant antibody construct and CD28 costimulation on lysis of malignant B cells from patients with chronic lymphocytic leukaemia by autologous T cells. Int J Cancer 112:509

    Article  PubMed  CAS  Google Scholar 

  • Ridgeway JB, Presta LG, Carter P (1996) “Knobs-into-holes” engineering of antibody CH3 domains for heavy chain heterodimerization. Protein Eng 9:617–621

    Article  Google Scholar 

  • Rohrbach F, Gerstmayer B, Biburger M, Wels W (2000) Construction and characterization of bispecific costimulatory molecules containing a minimized CD86 (B7-2) domain and single-chain antibody fragments for tumor targeting. Clin Cancer Res 6:4314–4322

    PubMed  CAS  Google Scholar 

  • Roopenian DC, Akilesh S (2007) FcRn: the neonatal Fc receptor comes of age. Nat Rev Immunol 7:715–725

    Article  PubMed  CAS  Google Scholar 

  • Roovers RC, Laeremans T, Huang L, De Taeye S, Verkleij AJ, Revets H, de Haard HJ, van Bergen en Henegouwen PMP (2007) Efficient inhibition of EGFR signalling and of tumor growth by antagonistic anti-EGFR nanobodies. Cancer Immunol Immunother 56:303–317

    Article  PubMed  CAS  Google Scholar 

  • Rossi EA, Goldenberg DM, Cardillo TM, McBride WJ, Sharkey RM, Chang CH (2006) Stably tethered multifunctional structures of defined composition made by the dock and lock method for use in cancer targeting. Proc Natl Acad Sci USA 103:6841–6846

    Article  PubMed  CAS  Google Scholar 

  • Ruf P, Lindhofer H (2001) Induction of a long-lasting antitumor immunity by a trifunctional bispecific antibody. Blood 98:2526–2534

    Article  PubMed  CAS  Google Scholar 

  • Russell JH, Ley TJ (2002) Lymphocyte-mediated cytotoxicity. Annu Rev Immunol 20:323–370

    Article  PubMed  CAS  Google Scholar 

  • Saerens D, Ghassabeh GH, Muyldersmans S (2008) Single-domain antibodies as building blocks for novel therapeutics. Curr Opin Pharmacol 8:600–608

    Article  PubMed  CAS  Google Scholar 

  • Schlereth B, Fichtner I, Lorenczewski G, Kleindienst P, Brischwein K, da Silva A, Kufer P, Lutterbüse R, Juhnhahn I, Kasimir-Bauer S, Wimberger P, Kimmig R, Baeuerle PA (2005) Eradication of tumors from a human colon cancer cell line and from ovarian cancer metastases in immunodeficient mice by a single-chain Ep-CAM-/CD3-bispecific antibody construct. Cancer Res 65:2882–2889

    Article  PubMed  CAS  Google Scholar 

  • Schneider MA, Bruhl H, Wechselberger A, Cihak J, Stangassinger M, Schlondorff D, Mack M (2005) In vitro and in vivo properties of a dimeric bispecific single-chain antibody IgG-fusion protein for depletion of CCR2(+) target cells in mice. Eur J Immunol 35:987–995

    Article  PubMed  CAS  Google Scholar 

  • Schoonjans R, Willems a, Schoonooghe S, Fiers W, Grooten J, Mertens N (2000) Fab chains as an efficient heterodimerization scaffold for the production of recombinant bispecific and trispecific antibody derivatives. J Immunol 165:7050–7057

    PubMed  CAS  Google Scholar 

  • Schroeder HW, Cavacini L (2010) Structure and function of immunoglobulins. J Allergy Clin Immunol 125:S41–S52

    Article  PubMed  Google Scholar 

  • Segal DM, Weiner GJ, Weiner LM (1999) Bispecific antibodies in cancer therapy. Curr Op Immunol 11:558–562

    Article  CAS  Google Scholar 

  • Shahied LS, Tang Y, Alpaugh RK, Somer R, Greenspon D, Weiner LM (2004) Bispecific minibodies targeting (HER2/neu and CD16 exhibit improved tumor lysis when placed in a divalent tumor antigen binding format. J Biol Chem 279:53907–53914

    Article  PubMed  CAS  Google Scholar 

  • Shalaby MR, Shepard HM, Presta L, Rodrgues ML, Beverley PCL, Feldmann M, Carter P (1992) Development of humanised bispecific antibodies reactive with cytotoxic lymphocytes and tumor cells overexpressing the HER2 protoncogene. J Exp Med 175:217–225

    Article  PubMed  CAS  Google Scholar 

  • Sharkey RM, Cardillo TM, Rossi EA, Chang CH, Karacay H, McBride WJ, Hansen HJ, Horak ID, Goldenberg DM (2005a) Signal amplification in molecular imaging by pretargeting a multivalent, bispecific antibody. Nat Med 11:1250–1255

    Article  PubMed  CAS  Google Scholar 

  • Sharkey RM, Karacay H, Chang CH, McBride WJ, Horak ID, Goldenberg DM (2005b) Improved therapy of non-Hodgkin’s lymphoma xenografts using radionuclides pretargeted with a new anti-CD20 bispecific antibody. Leukemia 19:1064–1069

    Article  PubMed  CAS  Google Scholar 

  • Skerra A (2007) Alternative non-antibody scaffolds for molecular recognition. Curr Opin Biotechnol 18:295–304

    Article  PubMed  CAS  Google Scholar 

  • Smith W, Jarrett AL, Beattie RE, Corvalan JR (1992) Immunoglobulins secreted by a hybrid-hybridoma: analysis of chain assemblies. Hybridoma 11:87–98

    Article  PubMed  CAS  Google Scholar 

  • Smith BJ, Popplewell A, Athwal D, Chapman AP, Heywood S, West SM, Carrington B, Nesbitt A, Lawson AD, Antoniw P, Eddelston A, Suittler A (2001) Prolonged in vivo residence times of antibody fragments associated with albumin. Bioconjug Chem 12:7450–7756

    Article  CAS  Google Scholar 

  • Stockmeyer B, Elsässer D, Dechant M, Repp R, Gramatzki M, Glennie MJ, van de Winkel JGJ, Valerius T (2001) Mechanisms of G-CSF- or GM-CSF-stimulated tumor cell killing by Fc receptor-directed bispecific antibodies. J Immunol Meth 248:103–111

    Article  CAS  Google Scholar 

  • Ternant D, Paintaud G (2005) Pharmacokinetics and concentration-effect relationships of therapeutic monoclonal antibodies and fusion proteins. Expert Opin Biol Ther 5:S37–S47

    Article  PubMed  CAS  Google Scholar 

  • Thakur A, Lum LG (2010) Cancer therapy with bispecific antibodies: clinical experience. Curr Opin Mol Ther 12:340–349

    PubMed  CAS  Google Scholar 

  • Tutt AL, Reid R, Wilkins BS, Glennie MJ (1995) Activation and preferential expansion of rat cytotoxic (CD8) T cells in vitro and in vivo with a bispecific (anti-TRC a/b x anti-CD2) F(ab’)2 antibody. J Immunol 155:2960–2971

    PubMed  CAS  Google Scholar 

  • Valerius T, Würflein D, Stockmeyer B, Repp R, Kalden JR, Gramatzki M (1997) Activated neutrophils as effector cells for bispecific antibodies. Cancer Immunol Immunother 45:142–145

    Article  PubMed  CAS  Google Scholar 

  • Vallera DA, Chen H, Sichender AR, Panoskaltsis-Mortari A, Taras EP (2009) Genetic alteration of a bispecific ligand-directed toxin targeting human CD19 and CD22 receptors resulting in improved efficacy against systemic B cell malignancies. Leuk Res 33:1233–1242

    Article  PubMed  CAS  Google Scholar 

  • Vallera DA, Oh S, Chen H, Shu Y, Frankel AE (2010) Bioengineering a unique deimmunized bispecific targeted toxin that simultaneously recognizes human CD22 and CD19 receptors in a mouse model of B-cell metastasis. Mol Cancer Ther 9:1872–1883

    Article  PubMed  CAS  Google Scholar 

  • van Spriel AB, van Ojik HH, van de Winkel JGJ (2000) Immunotherapeutic perspective for bispecific antibodies. Immunol Today 21:391–396

    Article  PubMed  Google Scholar 

  • Völkel T, Korn T, Bach M, Müller R, Kontermann RE (2001) Optimized linker sequences for the expression of monomeric and dimeric bispecific single-chain diabodies. Protein Eng 14:815–823

    Article  PubMed  Google Scholar 

  • Wallace PK, Kaufman PA, Lewis LD, Keler T, Givan AL, Fisher JL, Waugh MG, Wahner AE, Guyre PM, Fanger MW, Ernstoff MS (2001) Bispecific antibody-targeted phagocytosis of HER-2/neu expressing tumor cells by myeloid cells activated in vivo. J Immunol Meth 248:167–182

    Article  CAS  Google Scholar 

  • Warnaar SO, de Paus V, Lardenoije R, Machielse BN, de Graaf J, Bregonje M, van Haarlem H (1994) Purification of bispecific F(ab’)2 from murine trinoma OC/TR with specificity for CD3 and ovarian cancer. Hybridoma 13:519–526

    PubMed  CAS  Google Scholar 

  • Weiner LM, Clark JI, Davey M, Li WS, Garcia de Palazzo I, Ring DB, Alpaugh RK (1995) Phase I trial of 2B1, a bispecific monoclonal antibody targeting c-erbB-2 and FcγRIII. Cancer Res 55:4586–4593

    PubMed  CAS  Google Scholar 

  • Wolf E, Hofmeister R, Kufer P, Schlereth B, Baeuerle PA (2005) BiTEs: bispecific antibody constructs with unique anti-tumor activity. Drug Disov Today 10:1237–1244

    Article  CAS  Google Scholar 

  • Wozniak-Knopp G, Bartl S, Bauer A, Mostageer M, Woisetschläger M, Antes B, Ettl K, Kainer M, Weberhofer G, Wiederkum S, Himmler G, Mudde GC, Rüker F (2010) Introducing antigen-binding sites in structural loops of immunoglobulin constant domains: Fc fragments with engineered Her2/neu-binding sites and antibody properties. Protein Eng Des Sel 23:289–297

    Article  PubMed  CAS  Google Scholar 

  • Wu C, Ying H, Grinnell C, Bryant S, Miller R, Clabbers A, Bose S, McCarty D, Zhu R-R, Santora L, Davis-Taber R, Kunes Y, Fung E, Schwartz A, Sakorafas P, Gu J, Tarcsa E, Murtaza A, Ghayur T (2007) Simultaneous targeting of multiple disease mediators by a dual-variable-domain immunoglobulin. Nat Biotechnol 25:1290–1297

    Article  PubMed  CAS  Google Scholar 

  • Wu C, Ying H, Bose S, Miller R, Medina L, Santora L, Ghayur T (2009) Molecular construction and optimization of anti-human IL-1alpha/beta dual variable domain immunoglobulins (DVD-Ig) molecules. mAbs 1:339–347

    Article  PubMed  Google Scholar 

  • Xie Z, Shi M, Feng J, Yu M, Sun Y, Shen B, Guo N (2003) A trivalent anti-erbB2/anti-CD16 bispecific antibody retargeting NK cells against human breast cancer cells. Biochem Biophys Res Commun 311:307–312

    Article  PubMed  CAS  Google Scholar 

  • Xie Z, Guo N, Yu M, Hu M, Shen B (2005) A new format of bispecific antibody: highly efficient heterodimerization, expression and tumor cell lysis. J Immunol Meth 296:95–101

    Article  CAS  Google Scholar 

  • Zeidler R, Mysliwietz J, Csanady M, Walz A, Ziegler I, Schmitt B, Wollenberg B, Lindhofer H (2000) The Fc-region of a new class of intact bispecific antibody mediates activation of accessory cells and NK cells and induces direct phagocytosis of tumor cells. Br J Cancer 83:261–266

    Article  PubMed  CAS  Google Scholar 

  • Zeidler R, Mayer A, Gires O, Schmitt B, Mack BM, Lindhofer H, Wollenberg B, Walz A (2001) TNFa contributes to the antitumor activity of a bispecific, trifunctional antibody. Anticancer Res 21:3499–3503

    PubMed  CAS  Google Scholar 

  • Zhu Z, Presta LG, Zapata G, Carter P (1997) Remodeling domain interfaces to enhance heterodimer formation. Protein Sci 6:781–788

    Article  PubMed  CAS  Google Scholar 

  • Zuo Z, Jimenez X, Witte L, Zhu Z (2000) An efficient route to the production of an IgG-like bispecific antibody. Protein Eng 13:361–367

    Article  PubMed  CAS  Google Scholar 

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  1. Institut für Zellbiologie und Immunologie, Universität Stuttgart, Allmandring 31, 70569, Stuttgart, Germany

    Roland E. Kontermann

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Kontermann, R.E. (2011). Bispecific Antibodies: Developments and Current Perspectives. In: Kontermann, R. (eds) Bispecific Antibodies. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-20910-9_1

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