Enhancing antibody Fc heterodimer formation through electrostatic steering effects: applications to bispecific molecules and monovalent IgG.
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.
Gunasekaran
Kannan
K
Department of Protein Science, Amgen Inc, Seattle, Washington 98119, USA. kannan@amgen.com
Pentony
Martin
M
Shen
Min
M
Garrett
Logan
L
Forte
Carla
C
Woodward
Anne
A
Ng
Soo Bin
SB
Born
Teresa
T
Retter
Marc
M
Manchulenko
Kathy
K
Sweet
Heather
H
Foltz
Ian N
IN
Wittekind
Michael
M
Yan
Wei
W
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
D018033:Antibodies, Bispecific; D007074:Immunoglobulin G; D014409:Tumor Necrosis Factor-alpha
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