Wang Y.-L.,Guiyang Medical University |
Wang Y.-L.,Guiyang Medical College |
Liu T.,Guiyang Medical University |
Liu T.,Guiyang Medical College |
And 9 more authors.
Chinese Journal of New Drugs | Year: 2015
Objective: To investigate the expression and purification of a novel fusion protein, RANKL-Fc. Methods: The carboxyl-terminal 177 amino acids of RANKL (140~317) and the 232 amino acids of human IgG Fc (1~232) were amplified by PCR. The amplified DNA fragment was subcloned into a mammalian cell expression vector (PCDNA 3.1) to generate a fusion protein of RANKL-Fc for protein expression in mammalian CHO cells. The interactions of RANKL-Fc with RANKL and Fc RII were investigated by cell staining and flow cytometric analysis, respectively. Results and Conclusion: RANKL-Fc was successfully expressed and purified in the CHO-K1 cells with a purity of greater than 90%. Preliminary studies demonstrated that this fusion protein could interact with RANK and Fc RII. This result indicates that this fusion protein might work as an alternative agent to treat cancer metastasis by targeting RANKL/RANK pathway. ©, 2015, Chinese Journal of New Drugs Co. Ltd. All right reserved.