›› 2010, Vol. 37 ›› Issue (3): 171-174.

• 疾病防治 • 上一篇    下一篇

盐酸四环素人工抗原的合成及鉴定

张桂贤   

  1. (山东临沂师范学院生命科学学院, 临沂 276005)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-03-20 发布日期:2010-03-20

Synthesis and Identification of Artificial Antigen against Tetracycline Hydrochloride

ZHANG Gui-xian   

  1. (College of Life Science, Linyi Normal College, Linyi 276005,China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-03-20 Published:2010-03-20

摘要: 本试验旨在合成盐酸四环素人工抗原,为其单克隆抗体的制备奠定试验基础。将盐酸四环素(TCH)发生霍夫曼降解和重氮化反应生成重氮盐,再与蛋白质载体(BSA,OVA)偶联生成免疫原和包被原,对偶联产物采用三氯化铁及紫外扫描法鉴定偶联成功,且免疫原和包被原的摩尔结合比分别约为3.4∶1和4.2∶1。用免疫原免疫BALB/c小鼠,用间接ELISA检测抗体效价,采用竞争ELISA法测IC50,获得了效价为1∶51200、IC50为16.91 ng/mL的多克隆抗体,最终证明人工抗原制备成功。

关键词: 盐酸四环素; 霍夫曼; 重氮化; 偶联; 载体

Abstract: Hofmann degradation and diazo-reaction of Tetracycline hydrochloride (TCH) happened and conjugated to protein-carrier (BSA, OVA).The conjuagates were synthesized by the means of FeCl3 resolution and UV scanning spectra, and mol conjugation ration of TCH to BSA was 3.2∶1 and TCH to OVA was 4.2∶1.The immunogen(TCH-BSA) was immunized to BALB/c mice and polycolnal antibodies were obtained. Tiler of the antibodies were 1∶51200 by the indirect ELISA and IC50 was 16.91 ng/mL by the competitive indirect ELISA. It showed that the artificial-synthesized antigen had immunogenicity.

Key words: Tetracycline hydrochloride; Hofmann degradation; diazo-reaction; conjugation; carrier

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