摘要
以离子交换、疏水作用、HPLC和凝胶过滤层析等方法对山羊乳腺生物反应器表达的人促红细胞生成素 (erythropoietin ,EPO)进行了分离纯化 ,初步探讨了其分离工艺 ,结果得到了纯度达到 96%以上的纯品 ,所纯化的EPO分子质量为 32kDa ,糖基化程度根据理论分子质量测算为40 %。Western blot证明所得纯化产品具有天然EPO免疫原性 ,N端氨基酸序列测定与文献报道一致 ,ELISA检测免疫活性为 2 4 0 0 0 0IU mg,表明通过 4步纯化步骤可以得到高纯度的产品 。
EPO expressed in goat mammary gland bioreactor was purified with IEC,HIC,HPLC and SEC.Finally,the purity of the EPO was reached 96%,which had the same immunogenicity and N amino acid sequence as the natural product.It also has high specific activity using ELISA detection,so the purification process is very effective and it also provides valuable information for scaling up of industrial production.
出处
《中国生物工程杂志》
CAS
CSCD
2004年第8期59-62,共4页
China Biotechnology
基金
国家"8 63"计划资助项目 ( 2 0 0 2AA2 0 665 1)
