title: Enhancing antibody Fc heterodimer formation through electrostatic steering effects: applications to bispecific molecules and monovalent IgG.
abstract: Naturally occurring IgG antibodies are bivalent and monospecific. Bispecific antibodies having binding specificities for two different antigens can be produced using recombinant technologies and are projected to have broad clinical applications. However, co-expression of multiple light and heavy chains often leads to contaminants and pose purification challenges. In this work, we have modified the CH3 domain interface of the antibody Fc region with selected mutations so that the engineered Fc proteins preferentially form heterodimers. These novel mutations create altered charge polarity across the Fc dimer interface such that coexpression of electrostatically matched Fc chains support favorable attractive interactions thereby promoting desired Fc heterodimer formation, whereas unfavorable repulsive charge interactions suppress unwanted Fc homodimer formation. This new Fc heterodimer format was used to produce bispecific single chain antibody fusions and monovalent IgGs with minimal homodimer contaminants. The strategy proposed here demonstrates the feasibility of robust production of novel Fc-based heterodimeric molecules and hence broadens the scope of bispecific molecules for therapeutic applications.
authors.affiliation: Department of Protein Science, Amgen Inc, Seattle, Washington 98119, USA. kannan@amgen.com
authors.forename: Kannan
authors.initials: K
authors.lastname: Gunasekaran
authors.forename: Martin
authors.initials: M
authors.lastname: Pentony
authors.forename: Min
authors.initials: M
authors.lastname: Shen
authors.forename: Logan
authors.initials: L
authors.lastname: Garrett
authors.forename: Carla
authors.initials: C
authors.lastname: Forte
authors.forename: Anne
authors.initials: A
authors.lastname: Woodward
authors.forename: Soo Bin
authors.initials: SB
authors.lastname: Ng
authors.forename: Teresa
authors.initials: T
authors.lastname: Born
authors.forename: Marc
authors.initials: M
authors.lastname: Retter
authors.forename: Kathy
authors.initials: K
authors.lastname: Manchulenko
authors.forename: Heather
authors.initials: H
authors.lastname: Sweet
authors.forename: Ian N
authors.initials: IN
authors.lastname: Foltz
authors.forename: Michael
authors.initials: M
authors.lastname: Wittekind
authors.forename: Wei
authors.initials: W
authors.lastname: Yan
mesh_terms: D000818:Animals; D018033:Antibodies, Bispecific / Q000737:chemistry*; D019281:Dimerization; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry*; D007700:Kinetics; D007963:Leukocytes, Mononuclear / Q000166:cytology; D013058:Mass Spectrometry / Q000379:methods; D051379:Mice; D008810:Mice, Inbred C57BL; D016688:Mice, Inbred NOD; D009368:Neoplasm Transplantation; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D014409:Tumor Necrosis Factor-alpha / Q000737:chemistry
chemical_list: D018033:Antibodies, Bispecific; D007074:Immunoglobulin G; D014409:Tumor Necrosis Factor-alpha
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