【摘要】 利用酶联免疫测定法(ELISA)测定乳中孕酮含量来进行妊娠诊断的的方法有很多,采用二级竞争酶联免疫法可提高反应的灵敏度,但操作可进一步简化。通过制备抗孕酮Fab’可简化该操作。实验采用硫酸铵沉淀法对兔抗孕酮血清进行粗提,再用DEAESepharoseFastFlow对IgG作进一步纯化,回收率为93.24%。用胃蛋白酶将IgG消化成F(ab’)2,回收率为73.75%。F(ab’)2用10mmol/L巯基乙醇在pH6.0,37℃条件下反应2h而被还原成Fab’,回收率为67.84%。最后对IgG、F(ab’)2、Fab’进行SDS-聚丙酰胺凝胶电泳其分子量分别接近于150000、92000、46000。更多还原

【Abstract】 Ammonium sulfate and DEAE Sepharose Fast Flow were employed to isolate an d purify the antibody against 11-α-hydroxyprogesterone -11-hemisuccinate:BSA from rabbit serum.The purified IgG was digested with pepsin a t 37℃for24h.Separated F (ab’ )2was then reduced to Fab’ with 10mmol /Lβ-mercaptoenthanol(pH 6.0)for 2h at 37℃.This Fab’ against progesterone can be used to i mprove the sensitivity and simplify the procedure of a newly developed double competition ELISA for early pregnan cy diagnosis of dairy cows.更多还原