Sang Taek Jung and Tae Hyun Kang contributed equally to this work.
Efficient expression and purification of human aglycosylated Fcγ receptors in Escherichia coli†
Article first published online: 7 MAY 2010
Copyright © 2010 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 107, Issue 1, pages 21–30, 1 September 2010
How to Cite
Jung, S. T., Kang, T. H. and Georgiou, G. (2010), Efficient expression and purification of human aglycosylated Fcγ receptors in Escherichia coli. Biotechnol. Bioeng., 107: 21–30. doi: 10.1002/bit.22785
- Issue published online: 16 JUL 2010
- Article first published online: 7 MAY 2010
- Accepted manuscript online: 7 MAY 2010 12:00AM EST
- Manuscript Accepted: 27 APR 2010
- Manuscript Revised: 16 APR 2010
- Manuscript Received: 12 FEB 2010
- Clayton Foundation
- Norman Hackerman Advanced Research Program of the State of Texas
- Fc gamma receptor (FcγR);
- IgG binding;
- bacterial expression;
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. Biotechnol. Bioeng. 2010;107: 21–30. © 2010 Wiley Periodicals, Inc.