Engineered aglycosylated full-length IgG Fc variants exhibiting improved FcγRIIIa binding and tumor cell clearance

Migyeong Jo, Hyeong Sun Kwon, Kwang Hoon Lee, Ji Chul Lee, Sang Taek Jung

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

FcγRIIIa, which is predominantly expressed on the surface of natural killer cells, plays a key role in antibody-dependent cell-mediated cytotoxicity (ADCC), a major effector function of therapeutic IgG antibodies that results in the death of aberrant cells. Despite the potential uses of aglycosylated IgG antibodies, which can be easily produced in bacteria and do not have complicated glycan heterogeneity issues, they show negligible binding to FcγRIIIa and abolish the activation of immune leukocytes for tumor cell clearance, in sharp contrast to most glycosylated IgG antibodies used in the clinical setting. For directed evolution of aglycosylated Fc variants that bind to FcγRIIIa and, in turn, exert potent ADCC effector function, we randomized the aglycosylated Fc region of full-length IgG expressed on the inner membrane of Escherichia coli. Multiple rounds of high-throughput screening using flow cytometry facilitated the isolation of aglycosylated IgG Fc variants that exhibited higher binding affinity to FcγRIIIa-158V and FcγRIIIa-158F compared with clinical-grade trastuzumab (Herceptin®). The resulting aglycosylated trastuzumab IgG antibody Fc variants could elicit strong peripheral blood mononuclear cell-mediated ADCC without glycosylation in the Fc region.

Original languageEnglish
Pages (from-to)278-289
Number of pages12
JournalmAbs
Volume10
Issue number2
DOIs
Publication statusPublished - 2018 Feb 17
Externally publishedYes

Fingerprint

Antibody-Dependent Cell Cytotoxicity
Immunoglobulin G
Antibodies
Neoplasms
Glycosylation
Natural Killer Cells
Polysaccharides
Blood Cells
Flow Cytometry
Leukocytes
Cell Death
Escherichia coli
Bacteria
Membranes
Trastuzumab
Therapeutics

Keywords

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

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Engineered aglycosylated full-length IgG Fc variants exhibiting improved FcγRIIIa binding and tumor cell clearance. / Jo, Migyeong; Kwon, Hyeong Sun; Lee, Kwang Hoon; Lee, Ji Chul; Jung, Sang Taek.

In: mAbs, Vol. 10, No. 2, 17.02.2018, p. 278-289.

Research output: Contribution to journalArticle

Jo, Migyeong ; Kwon, Hyeong Sun ; Lee, Kwang Hoon ; Lee, Ji Chul ; Jung, Sang Taek. / Engineered aglycosylated full-length IgG Fc variants exhibiting improved FcγRIIIa binding and tumor cell clearance. In: mAbs. 2018 ; Vol. 10, No. 2. pp. 278-289.
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