2,4-D人工抗原合成及抗体制备_生物工程.doc

摘要:2,4-D是苯氧羧酸类除草剂，为半抗原。由于2,4-D农药残留危害着生态环境和人类的健康，所以对2,4-D的残留检测已经越来越重要。以牛血清白蛋白(BSA)和卵清白蛋白(OVA)为载体蛋白，采用碳化二亚胺法(EDC法)，将2,4-D分别与BSA和OVA偶联，合成免疫抗原2,4-D-BSA和包被抗原2,4-D-OVA。采用紫外扫描(UV)进行鉴定，结果表明，2,4-D与BSA偶联后，其波峰为278nm，与BSA、2,4-D扫面后的波峰相比，出现了明显的变化，由此证明半抗原与载体蛋白偶联成功。用合成的免疫抗原2,4-D-BSA按常规方法免疫小鼠，经间接ELISA法检测小鼠血清效价，效价比均在1:6400以上，证明免疫小鼠产生了抗体。这为2,4-D的免疫分析检测提供了理论与实际操作的经验。

关键词：2,4-D；人工抗原；偶联；抗体

Abstract:2,4-D is a kind of phenoxy acid herbicide and hapten.Because the pesticide residue of 2,4-D does harm to ecological environment and human health, so the residue detection of 2,4-D has been more and more important.Using the bovine serum albumin(BSA) and the ovalbumin(OVA) as a carrier protein,adapting the method of EDC,coupling 2,4-D with BSA and OVA respectively, in order to synthesis the immunizing antigen that it is 2,4-D-BSA and the envelope antigen that it is 2,4-D-OVA. Results show that by using ultraviolet scanning (UV) to identify,coupled with the 2,4-D and BSA,the peak is 278nm.It compares with the peak of BSA and 2,4-D,the peak appears obvious changes,so the carrier protein is successfully coupled.By conventional method of immune mice with the immunizing antigen.Using ELISA to detect the serum titer of mice.The potency ratio is above 1:6400.It proves immune mice develop antibodies.By way of providing theory and practical experience for the  immunoassay of 2,4-D.

Keywords  2,4-D   Artificial antigen   Coupling   Antibody