重组人干细胞因子理化性质和生物活性分析

Physico-chemical property and bioactivity determination of recombinant stem cell factor

干细胞因子 (stemcellfactor ,SCF)是一种对造血细胞有重要作用的因子 ,在自体外周血造血前体细胞移植、肿瘤放化疗的辅助治疗、贫血和其他血液病的治疗方面有良好的应用前景。目的 :分析本所制备的rhSCF的理化性质和生物活性。方法 :用反相高效液相色谱、质谱、凝胶高效液相色谱等方法分别测定了制品的纯度、分子量、氨基酸组成、紫外光谱等 ,并用TF 1细胞测定了制品的生物活性。结论 :制品分子量为 18558,与天然hSCF的差异小于 0 .2 % ,产物以非共价二聚体形式存在。氨基酸组成与天然hSCF相同 ,紫外光谱为典型的蛋白光谱 ,在 2 77nm有特征吸收 ,比活为 0 .95× 10 6U /mg ,与国外参考品相似。上述结果初步说明制备的rhSCF理化性质与天然hSCF一致 ,生物活性良好 ,有望进一步开发用于临床。

Stem cell factor(SCF) is an important hematopoietic growth factor,it may be useful in mobilization of autologous peripheral blood progenitor cells for transplantation, supplementary therapy of tumor,therapy of anemia and other hematopoietic diseases. Objective:To determine the physico-chemical property and bioactivity of recombinant stem cell factor produced by our institute. Methods: The purity, molecular weight, amino acid composition and UV spectra of product were determined by RP-HPLC , MS, gel filtration-HPLC etc. The bioactivity of product was determined by TF-1 cell.Conclusions:The purity of product was greater than 98% and the molecular weight determined was 18 558 which was different from natural hSCF by less than 0.2%. The product was an uncovalent dimer.The results of amino acid composition and N-terminal amino acid sequence analysis were similar to those of natural hSCF except having an additional methionine.The UV spectrum was an typical protein one with special absorption at 277 nm.The specific activity of rhSCF was 0.95×10 6 U/mg,which was similar to the standard of NIBSC. All these results showed the physico-chemical property of product was similar to the natural one.It has high biological activity, and may be useful in hematopathy. [