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 language | English |
|---|---|
| Pages (from-to) | 21-30 |
| Number of pages | 10 |
| Journal | Biotechnology and Bioengineering |
| Volume | 107 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 2010 Sep 1 |
| Externally published | Yes |
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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 journal › Article
}
TY - JOUR
T1 - Efficient expression and purification of human aglycosylated Fcγ receptors in Escherichia coli
AU - Jung, Sang Taek
AU - Kang, Tae Hyun
AU - Georgiou, George
PY - 2010/9/1
Y1 - 2010/9/1
N2 - 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.
AB - 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.
KW - Bacterial expression
KW - Fc gamma receptor (FcγR)
KW - Glycosylation
KW - IgG binding
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UR - http://www.scopus.com/inward/citedby.url?scp=77956260081&partnerID=8YFLogxK
U2 - 10.1002/bit.22785
DO - 10.1002/bit.22785
M3 - Article
C2 - 20506277
AN - SCOPUS:77956260081
VL - 107
SP - 21
EP - 30
JO - Biotechnology and Bioengineering
JF - Biotechnology and Bioengineering
SN - 0006-3592
IS - 1
ER -