抗孕酮Fab’-大豆过氧化物酶交联物的制备和鉴定  

PREPARATION AND IDENTIFICATION OF ANTI-PROGESTERONE Fab'-SBP

作　　者：龚振明[1] 李蓓[1] 陈鲁勇[1] 沈明泉[1] 

机构地区：[1]上海交通大学农业与生物学院,上海201101

出　　处：《上海农业学报》2003年第1期76-79,共4页Acta Agriculturae Shanghai

基　　金：上海市高校科技发展基金 (98E0 7)资助

摘　　要：用硫酸铵沉淀法对兔抗孕酮血清进行粗提后用DEAESepharoseFastFlow对IgG进行层析纯化 ,然后用胃蛋白酶切除Fc片段而制备F(ab’) 2 ,用 β 巯基乙醇还原F(ab’) 2 后得到Fab’ ,再将Fab’与热稳定性较高的大豆过氧化物酶 (SBP)偶联制得抗孕酮Fab’ SBP。将抗孕酮Fab’ SBP用于二级竞争酶联免疫法测定孕酮有以下优点 :(1)减少因Fc片段与固相载体非特异性吸附而引起的高背景干扰 ;(2 )由于Fab’和SBP在很温和的条件下偶联 。Rabbit anti progesterone IgG, prepared by ammonium sulfate precipitation and DEAE Sepharose Fast Flow chromatography purification, was digested with pepsin. F(ab') 2, one of the enzyme digestion products, was isolated with Sephadex 150 and reduced into Fab' by β mercaptoethanol. The Fab' was then coupled with SBP Mal (Activated Soybean Peroxidase EZ Link TM Maleimide) to produce Fab' SBP for a double competition EIA progesterone measurement strategy. The purpose of this work was to (1) minimize the background interference caused by non specific adsorption between Fc and solid phase carrier, (2) maximize the immuno activity and enzyme activity of Fab' SBP by coupling the reagents in a mild condition.

关 键 词：抗孕酮Fob’-SBP 大豆过氧化物酶 交联物 制备 鉴定 酶联免疫测定 

分 类 号：S852.4[农业科学—基础兽医学]