抗体夹心酶免疫吸附法测定重组E.coli水蛭素HV3研究  

Triple Antibodies Sandwich Enzyme-Linked Immunoadsorbent Assay for Determination of Recombinant E.coli Hirudin Ⅲ

作　　者：郭丰[1] 谭树华[1] 吴梧桐[1] 

机构地区：[1]中国药科大学生命科学与技术学院,江苏南京210009

出　　处：《药物生物技术》2006年第2期87-90,共4页Pharmaceutical Biotechnology

基　　金：国家教育部博士学科点基金(20040316005)

摘　　要：将重组E. coli水蛭素HV3和BSA交联后免疫豚鼠和家兔,DEAE-纤维素柱层析纯化IgG.用这两种抗体和酶标二抗羊抗兔IgG-HRP建立三抗体夹心酶联免疫吸附法,测定重组E. coli水蛭素HV3的含量.本测定方法的线性范围为0～2 ng/ml,灵敏度为0.04 ng/ml.建立的三抗体夹心酶联免疫吸附法测定重组E.coli水蛭素HV3具有良好的精密度、回收率、灵敏度和特异性.The purpose is to establish a ELISA method of determination for recombinent E. coli hirudin Ⅲ. Conjugation of recombinant E. coli hirudin Ⅲ to bovine serum albumin（BSA） was obtained using EDC method. A DHP guinea pig and a Chinese rabbit were immunized with recombinant E. coli hirudin Ⅲ BSA. The immunoglobulin G was seperated and purified by using DEAE cellulose DE 52 chromatography. Recombinant E. coli hirudin Ⅲ was measured by triple antibodies sandwich enzyme-linked immunoadsorhent assay using antibodies from guinea pig and rabhit and a horseradish peroxidase coniugated antibody against rabbit IgG from goat. The linearities and sensitivity were 0-2 ng/ml and 0. 04 ng/ml respectively. Triple antibodies sandwich enzyme-linked immunoadsorbent assay was constant, reliable, sensitive,and suitable for the determination of recombinant E. coli hirudin Ⅲ.

关 键 词：重组E.coli水蛭素HV3(rHV3) 交联 三抗体夹心酶联免疫吸附法 

分 类 号：Q51[生物学—生物化学]