Oxidation of M252 but not M428 in hu-IgG1 is responsible for decreased binding to and activation of hu-FcγRIIa (His131). (November 2017)

• Record Type: Journal Article
• Authors: Cymer, Florian; Thomann, Marco; Wegele, Harald; Avenal, Cecile; Schlothauer, Tilman; Gygax, Daniel; Beck, Hermann
• Is Part Of: Biologicals. Volume 50(2017)
• Page Start: 125
• Page End: 128
• Publication Date: 2017-11
• Subjects: Fc gamma receptor -- Antibody -- Oxidation -- Affinity chromatography -- ADCP; ADCP antibody dependent cellular phagocytosis -- CQA critical quality attribute -- FcγRIIa Fc gamma receptor IIa -- FcRn neonatal Fc-receptor -- H histidine -- M methionine -- R arginine -- mAb monoclonal antibody -- QbD quality by design -- SPR surface plasmon resonance; Biological products -- Standards -- Periodicals; Biological Products -- Periodicals; Biological Products -- standards -- Periodicals; Produits biologiques -- Normes -- Périodiques; Biological products -- Standards; Periodicals; 615.37
• DOI: 10.1016/j.biologicals.2017.09.006
• Languages: English
• ISSNs: 1045-1056

Abstract

Abstract: Oxidation of monoclonal therapeutic antibodies (mAbs) can affect binding to Fc-receptors and potentially influence pharmacokinetics or effector functions like e.g. antibody dependent cellular phagocytosis (ADCP). Recently, it has been demonstrated that binding to FcγRIIa (H131) is affected by methionine oxidation of the Fc-portion but it is currently unknown which methionine is responsible for decreased binding. We separated an oxidized IgG1 monoclonal antibody based on the oxidation state of methionine 252 and analyzed fractionated material in receptor binding experiments as well as in functional (cell-based) assays. Although the unfractionated mixture demonstrated weaker interaction/activation of the receptor, differently oxidized isolated subspecies can lead both to stronger as well as weaker binding and activation of the histidine variant of FcγRIIa.