title: Enhancement of antibody-dependent cell-mediated cytotoxicity by endowing IgG with FcαRI (CD89) binding.
abstract: Fc effector functions such as antibody-dependent cell-mediated cytotoxicity (ADCC) and antibody-dependent cell-mediated phagocytosis (ADCP) are crucial to the efficacy of many antibody therapeutics. In addition to IgG, antibodies of the IgA isotype can also promote cell killing through engagement of myeloid lineage cells via interactions between the IgA-Fc and FcαRI (CD89). Herein, we describe a unique, tandem IgG1/IgA2 antibody format in the context of a trastuzumab variable domain that exhibits enhanced ADCC and ADCP capabilities. The IgG1/IgA2 tandem Fc format retains IgG1 FcγR binding as well as FcRn-mediated serum persistence, yet is augmented with myeloid cell-mediated effector functions via FcαRI/IgA Fc interactions. In this work, we demonstrate anti-human epidermal growth factor receptor-2 antibodies with the unique tandem IgG1/IgA2 Fc can better recruit and engage cytotoxic polymorphonuclear (PMN) cells than either the parental IgG1 or IgA2. Pharmacokinetics of IgG1/IgA2 in BALB/c mice are similar to the parental IgG, and far surpass the poor serum persistence of IgA2. The IgG1/IgA2 format is expressed at similar levels and with similar thermal stability to IgG1, and can be purified via standard protein A chromatography. The tandem IgG1/IgA2 format could potentially augment IgG-based immunotherapeutics with enhanced PMN-mediated cytotoxicity while avoiding many of the problems associated with developing IgAs.
authors.affiliation: a Antibody Discovery and Protein Engineering; Medimmune Ltd. ; Gaithersburg , MD , USA.
authors.forename: M Jack
authors.initials: MJ
authors.lastname: Borrok
authors.forename: Nadia M
authors.initials: NM
authors.lastname: Luheshi
authors.forename: Nurten
authors.initials: N
authors.lastname: Beyaz
authors.forename: Gareth C
authors.initials: GC
authors.lastname: Davies
authors.forename: James W
authors.initials: JW
authors.lastname: Legg
authors.forename: Herren
authors.initials: H
authors.lastname: Wu
authors.forename: William F
authors.initials: WF
authors.lastname: Dall'Acqua
authors.forename: Ping
authors.initials: P
authors.lastname: Tsui
mesh_terms: D000818:Animals; D000920:Antibody-Dependent Cell Cytotoxicity / Q000187:drug effects* / Q000276:immunology; D015703:Antigens, CD / Q000276:immunology*; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D007070:Immunoglobulin A / Q000276:immunology / Q000494:pharmacology; D007074:Immunoglobulin G / Q000276:immunology / Q000494:pharmacology; D007962:Leukocytes / Q000276:immunology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010587:Phagocytosis / Q000187:drug effects*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000276:immunology; D011961:Receptors, Fc / Q000276:immunology*; D017452:Receptors, IgG / Q000276:immunology
chemical_list: D015703:Antigens, CD; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D018719:Erb-b2 Receptor Tyrosine Kinases; D011961:Receptors, Fc; D017452:Receptors, IgG; C036832:Fc(alpha) receptor; C508053:ERBB2 protein, human
keywords: ADCC; ADCC, antibody-dependent cell-mediated cytotoxicity; ADCP, antibody-dependent cell-mediated phagocytosis; AUC, area under the curve; CL, clearance rate; CD89; CDC, complement dependent cytotoxicity; Cmax, maximum serum concentration; DSC, differential scanning calorimetry; E:T ratio, effector to target ratio; FCM, flow cytometry; FcRn, neonatal Fc receptor; FcαRI; FcγR, Fc gamma receptor; HER2, human epithelial receptor two; IgA; IgA, immunoglobulin A; IgG, immunoglobulin G; LDH, lactate dehydrogenase; MΦ, macrophage; NK, natural killer; PBMC, peripheral blood mononuclear cell; PK, pharmacokinetics; PMN, polymorphonuclear; SPR, surface plasmon resonance; TAA, tumor associated antigens; T½, half-life; Vss, central compartment volume of distribution; macrophage; monoclonal antibody; neutrophil; tandem; trastuzumab