Overproduction and biological activities of recombinant human interleukin 6 expressed in E. coli = 대장균에서 재조합 인간 interleukin 6의 대량생산 및 그 생물학적 활성의 측정
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- Overproduction and biological activities of recombinant human interleukin 6 expressed in E. coli = 대장균에서 재조합 인간 interleukin 6의 대량생산 및 그 생물학적 활성의 측정
- Young Yang; Hyung Sik Kang; Woo Jin Na; Choong Eun Lee; Kwang Ho Pyun
- Bibliographic Citation
- Korean Biochemical Journal, vol. 25, no. 2, pp. 165-170
- Publication Year
- Human interleukin 6 (hIL-6) is a multifunctional cytokine involved in acute phase and immune response. While a critical role of IL-6 in various immune disorders has been suggested, studies on its detailed functional mechanism are often hampered due to the low natural abundance of this cytokine. Thus，for a large scale production of hIL-6, we have made an attempt to directly express hIL-6 using pET-8c expression plasmid under the control of T7 promoter in Escherichia coli, A cDNA coding for hIL-6 without the signal sequence was amplified using PCR reaction, fused to pET-8cf and transformed into E. coli (XDE3). Upon induction with isopropyl thiogalactoside, recombinant human interleukin 6 (rhIL-6) was overexpressed as a form of inculsion bodies, with its expression level reaching over 30% of total E. coli proteins. The rhIL-6 was isolated from inclusion bodies by solubilization in 6 M guanidine hydrochloride followed by dialysis against 50 mM Tris buffer. A single passage over DEAE-Sepharose faciliated the purification of rhIL-6, The purified rhIL-6 was characterized by nucleotide sequence analysis and bioassays. The biological activity was confirmed by proliferation of B9 cells and immunoglobulin secretion of SAC-blast B cells.
- Korea Soc-Assoc-Inst
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- 1. Journal Articles > Journal Articles
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