应用免疫印迹法检测人基因工程干扰素中大肠杆菌残余成分

用大肠杆菌抽提物免疫家兔制备相应的抗血清,并用该血清建立免疫印迹法,检测基因工程干扰素中大肠杆菌残余成分。初步试验结果表明,该方法的灵敏度高于SDS-PAGE银染法。检测3批人α_1.型基因工程干扰素的结果表明,制品的纯度较好。

An Immunobl otting Assay for the Detection of E. Coli Contaminants in Recombinant IFN Preparation

An i mmunoblotting assay was established to detect the E. coli contam inants in the final products of reeombinant IFN. Antiserum against E. coli cell components was prepared by immunizing rabbits with lysate of E. coli harboring no human IFN gene. In order to get a satisfactory composition of antibodies against all of the E. coli cell components, antisera from four rabbits were pooled before use. When lysate of E. coli was separated by SDS-PAGE and then i mmunoblotted with the antiE. coli antiserum, all of its components, which were detectable by silver staining method, were also clearly shown in the result. Further study indicated that the immunoblotting method was not only highly, specific, but also more sensitive than silver staining method (SDS-PAGE) in detecting the E. coli contami nants. By using this method, no E. coli contaminant was found in the final products of human recombinant IFN-α_(?) prepared in our institute.