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中国农学通报

中国农学通报 ›› 2010, Vol. 26 ›› Issue (6): 43-46.

所属专题： 生物技术

• 生物技术科学 • 上一篇    下一篇

人工合成氯霉素抗原免疫血清效价的比较及多克隆抗体的制备

张红星,韩景丽,陈静文,黄漫青,张馨如   

  • 收稿日期:2009-10-26 修回日期:2009-11-18 出版日期:2010-03-20 发布日期:2010-03-20
  • 基金资助:

    北京农学院青年科学基金

Comparison of titer of anti-sera from CAP-BSA 、CAP-OVA and Preparation of the polyclonal antibody against chloramphenicol

  • Received:2009-10-26 Revised:2009-11-18 Online:2010-03-20 Published:2010-03-20

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摘要：

摘要：【研究目的】人工合成氯霉素抗原及氯霉素多克隆抗体的制备，比较氯霉素人工抗原免疫血清效价。【方法】采用重氮化法，分别以BSA、OVA为载体蛋白合成CAP-BSA,CAP-OVA两种完全抗原，并对两种合成抗原的免疫效果进行比较。分别以这两种抗原作为免疫抗原用弗氏佐剂乳化后免疫新西兰大白兔，免疫四次后，心脏采血，分离血清，再用两种抗原分别作为包被原对兔血清进行间接竞争ELISA分析。【结果】大白兔经免疫后产生了抗氯霉素特异性抗体，两种抗原得到的抗体效价均达到1：8100，氯霉素最低检出限为9ng/ml。【结论】综合比较认为，CAP-OVA作为免疫原，CAP-BSA用作包被原是比较理想的实验方案，可用于制备氯霉素抗体和研制生产氯霉素酶联免疫试剂盒。

关键词: 大菱鲆, 大菱鲆, 豆粕, 发酵, 米曲霉

Abstract:

Abstract:【OBJECTIVE】Synthesis of CAP-BSA,CAP-OVA, polyclonal antibody against chloramphenicol, and comparison of titer of anti-sera from CAP-BSA,CAP-OVA.【METHOD】Bovine serum albumin (BSA) and Ovalbumin (OVA) were used as protein carries respectively to couple with chloramphenicol（CAP）by diazotization method, then compared the immune ability of the two synthetic antigens. The two synthesized antigens CAP-BSA and CAP-OVA were mixed with the same amount of adjuvant and then immunized rabbits. After the fourth immunization, the rabbits were blooded from heart, and then separated blood sera; lastly we issued indirect competitive ELISA analysis using the two antigens as coatingen respectively.【RESULTS】Enzyme linked immuno-sorbent assay (ELISA) indicates the existence of specific antibodies in the anti-sera. The titer of anti-sera for CAP-BSA and CAP-OVA is 1:8100. The detection limit for CAP was 9ng/mL.【CONCLUSION】After comparison of the results comprehensively, we believed that using CAP-OVA as immunogen and CAP-BSA as coatingen is a good experimental program. This experimental program can be used for preparation of polyclonal antibody against chloramphenicol and production of chloramphenicol ELISA Kit.