| 초록 |
Human Fc receptors are expressed as membrane-bound molecules on a variety of immune cells, and which are play well-defined role in regulation of innate and adaptive immune responses by binding to immunoglobulin G (IgG) class antibody. In this research work, we achieved large scale production of recombinant FcγRIIa by high cell density cultivation of E. coli. FcγRIIa expression was induced at two different cell concentrations, OD600 of 46 and 100. When E. coli cells were induced at higher cell concentration (OD600 of 100), cell concentration reached to OD600 of 234 in 9 h cultivation after induction. FcγRIIa was produced as insoluble inclusion body in cytoplasm and receptor productions were also maintained at high levels during induction period. The maximum content of FcγRIIa was 17.4% of total proteins at 10 h after induction and volumetric productivity was about 0.32 g/L/h.By simple purification procedures including denaturation and refolding process, 87 mg of soluble FcγRIIa could be obtained from 20 mL of culture with high purity.The biological activity of refolded FcγRIIa was also evaluated with its interaction to all subclasses of IgG antibodies. |
