Expression of truncated fragment of human OPG in CHO-DHFR- cells and its bioactivity characterization
10.3321/j.issn:1007-8738.2009.11.011
- VernacularTitle:截短型人骨保护素在CHO-DHFR-细胞中的表达及活性测定
- Author:
Bohong KANG
;
Xiaoyi ZANG
;
Lanying LI
;
Kongyin ZHAO
- Publication Type:Journal Article
- Keywords:
osteoprotegerin;
CHO-DHFR~- cell;
expression;
bioactivity analysis
- From:
Journal of Cellular and Molecular Immunology
2009;25(11):1002-1004
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To obtain high level expression of recombinant human truncated osteoprotegerin (TOPG) with higher bioactivity in CHO-DHFR~-cells. METHODS: The recombinant vector pcDNA3.1/DHFR-TOPG was constructed and transfected into CHO-DHFR~- Cells by the directions of LipofectAMINE~(TM)2000 for stable expression. The stable expression cell strains were screened by selective medium IMDM with 50 mL/L FCS, then serially passed in methotraxate (MTX) for gene amplification. The expression were analyzed by ELISA and RT-PCR. At last, the bioactivity analysis was performed in vitro. RESULTS: The expression level of recombinant truncated human OPG was up to 6 mg/L·72 h, and it had significant suppression effect on the formation of OLC(P<0.05). CONCLUSION: Recombinant truncated human OPG has high expression and bioactivity. The results make it possible for further studying and clinical implying of OPG.
