Efficient expression and purification of human aglycosylated Fcγ receptors in Escherichia coli

Sang Taek Jung, Tae Hyun Kang, George Georgiou

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Effector Fc gamma receptors (FcγRs) are expressed on the surface of a variety of cells of hematopoietic lineage and serve as a bridge between adaptive and innate immune responses. The interaction between immune complexes, formed by IgG class antibodies that are crosslinked with antigen, and FcγRs triggers signaling cascades that result in numerous cellular responses including the activation or donwregulation of cytotoxic responses, cytokine release, and antibody synthesis. Here, the extracellular domains of the human type I transmembrane FcγRs were expressed in Escherichia coli and their interactions to subclass IgGs (IgG1, IgG2, IgG3, and IgG4) antibodies were analyzed. Expression using fully synthetic E. coli codon optimized FcγR genes and optimization of sequences for N-terminal translation initiation region through mRNA secondary structure prediction enabled us to achieve high yield of purified, bacterially expressed receptors, including FcγRI and FcγRIIIa which have not been successfully expressed in bacteria until now. The aglycosylated FcγRs showed similar IgG subclass binding selectivity compared to the respective glycosylated FcγRs expressed in mammalian cells.

Original languageEnglish
Pages (from-to)21-30
Number of pages10
JournalBiotechnology and Bioengineering
Volume107
Issue number1
DOIs
Publication statusPublished - 2010 Sep 1
Externally publishedYes

Fingerprint

Fc Receptors
IgG Receptors
Antibodies
Escherichia coli
Purification
Immunoglobulin G
Antigens
Bacteria
Genes
Chemical activation
Cells
Immunoglobulin Isotypes
Adaptive Immunity
Cell Lineage
Antigen-Antibody Complex
Innate Immunity
Codon
Cytokines
Messenger RNA

Keywords

  • Bacterial expression
  • Fc gamma receptor (FcγR)
  • Glycosylation
  • IgG binding

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Efficient expression and purification of human aglycosylated Fcγ receptors in Escherichia coli. / Jung, Sang Taek; Kang, Tae Hyun; Georgiou, George.

In: Biotechnology and Bioengineering, Vol. 107, No. 1, 01.09.2010, p. 21-30.

Research output: Contribution to journalArticle

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