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∶51200I、C50为16.91 ng/mL的多克隆抗体,最终证明人工抗原制备成功。
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 FeCl3 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.
出处
《中国畜牧兽医》
CAS
北大核心
2010年第3期171-174,共4页
China Animal Husbandry & Veterinary Medicine
关键词
盐酸四环素
霍夫曼
重氮化
偶联
载体
Tetracycline hydrochloride
Hofmann degradation
diazo-reaction
conjugation
carrier
