重组人长效促卵泡激素FSH-CTP的纯化工艺研究及鉴定

Research and Appraisal on Purification Process of Recombinant Human Long-acting Follicle Stimulating Hormone

为了获得高纯度的重组人长效促卵泡激素FSH-CTP,收集表达FSH-CTP的CHO细胞培养上清,经过染料亲和层析、DEAE阴离子交换层析以及疏水层析三步层析制备,所得样品进行分子量、等电点测定和Western Blot鉴定,雌性大鼠卵巢增重法测定体内生物活性。结果显示：经3步纯化所得样品纯度可达98%以上,表观分子量约为47 k Da,MALDI-TOF-MS测得结果为35.2 k Da,等电点4.0-2.8,Western Blot结果呈阳性,测活结果表明,FSH-CTP的活性比重组人卵泡刺激素明显偏高。纯化所得样品纯度良好,为该药物的进一步研究打下了基础。

To obtain high purity recombinant human long-acting follicle stimulating hormone FSH-CTP,the study collected the CHO cells culture supernatant contained FSH-CTP,then it was puried by dye affinity chromatography,DEAE anion exchange chromatography and hydrophobic interaction chromatography. We detected the puried samples by molecular weight,isoelectric point,biological activity in vitro（ Western Blot） and biological activity in vivo（ ovarian weight gain in female rats law）. The results showed that： The purity of FSH-CTP protein could reach 98% after three stept purification,the apparent molecular weight was about 47 k Da,MALDI-TOF-MS result was 35.2 k Da. The results of isoelectric point was 4.0 ~ 2.8. And we provided the FSHCTP activity was higher than recombinant human follicle stimulating hormone significantly by Western Blot analysis. The purification and activity of FSH-CTP was high. The results could lay the foundation for further study of the drug.