Optimal combination of beneficial mutations for improved ADCC effector function of aglycosylated antibodies

Hyun Woung Yoon, Migyeong Jo, Sanghwan Ko, Hyeong Sun Kwon, Chung Su Lim, Byoung Joon Ko, Ji Chul Lee, Sang Taek Jung

Research output: Contribution to journalArticle

Abstract

The Fc region of IgG antibodies is crucial for binding to Fc receptors expressed on the surfaces of various immune leukocytes and eliciting therapeutic effector functions such as clearance of antibody-opsonized tumor cells. Despite abrogated Fc gamma receptor (FcγR) binding and therapeutic effector function in the absence of N-linked glycosylation at Asn297, the aglycosylated Fc region of IgG antibodies has bioprocessing advantages such as the absence of glycan heterogeneity and simple bacterial antibody production. Therefore, these antibodies have been comprehensively engineered as effector functional units for human therapy. In this work, we constructed a huge library of Fc variants with combinations of 25 beneficial mutations that were previously identified to improve binding of glycosylated or aglycosylated Fc regions to human FcγRs in previous studies. High-throughput screening of the resulting library led to the identification of an aglycosylated Fc variant that exhibited almost double the antibody-dependent cell-mediated cytotoxicity than wild-type glycosylated Fc. All mutations in this aglycosylated Fc variant were derived from previously identified beneficial mutations for engineered aglycosylated Fc variants as opposed to glycosylated variants, suggesting that significantly different sets of beneficial mutations are necessary to improve the effector function of aglycosylated Fc.

Original languageEnglish
Pages (from-to)62-71
Number of pages10
JournalMolecular Immunology
Volume114
DOIs
Publication statusPublished - 2019 Oct 1

Fingerprint

Antibody-Dependent Cell Cytotoxicity
Mutation
Antibodies
Bacterial Antibodies
Immunoglobulin G
Neoplasm Antibodies
IgG Receptors
Fc Receptors
Glycosylation
Libraries
Antibody Formation
Polysaccharides
Leukocytes
Therapeutics

Keywords

  • Aglycosylated IgG
  • Antibody-dependent cell-mediated cytotoxicity
  • Effector functions
  • Fc engineering
  • FcγRIIIa

ASJC Scopus subject areas

  • Immunology
  • Molecular Biology

Cite this

Optimal combination of beneficial mutations for improved ADCC effector function of aglycosylated antibodies. / Yoon, Hyun Woung; Jo, Migyeong; Ko, Sanghwan; Kwon, Hyeong Sun; Lim, Chung Su; Ko, Byoung Joon; Lee, Ji Chul; Jung, Sang Taek.

In: Molecular Immunology, Vol. 114, 01.10.2019, p. 62-71.

Research output: Contribution to journalArticle

Yoon, Hyun Woung ; Jo, Migyeong ; Ko, Sanghwan ; Kwon, Hyeong Sun ; Lim, Chung Su ; Ko, Byoung Joon ; Lee, Ji Chul ; Jung, Sang Taek. / Optimal combination of beneficial mutations for improved ADCC effector function of aglycosylated antibodies. In: Molecular Immunology. 2019 ; Vol. 114. pp. 62-71.
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