抗氯霉素单链抗体基因表达及免疫学活性检测

Expression and immunological activity of single-chain antibody against chloramphenicol

旨在构建和原核表达抗氯霉素的单链抗体（Sc Fv）基因,并进行免疫学活性的初步测定。将前期克隆的氯霉素Sc Fv全长基因克隆到p ET-24a载体后转化入大肠杆菌BL21,IPTG诱导后进行SDS-PAGE检测目的蛋白的表达,间接ELISA分析纯化的表达产物的免疫活性。结果：经酶切鉴定及DNA序列测定证明原核表达载体构建正确,SDS-PAGE分析显示表达蛋白为27.5 ku,间接ELISA表明纯化后的蛋白免疫学活性良好。本试验成功表达了具有免疫学活性的氯霉素Sc Fv,为氯霉素的药物残留检测方法建立奠定了基础。

This study was to construct a gene of ScFv against chloramphenlcol and to determine the immunological activity of the gene. The full-length clone of the Chloramphenicol ScFv gene was further cloned into a pET-24a vector and then transformed into E. coli BL21. The ex- pression of the target protein was induced by IPTG and was then detected by SDS-PAGE, and the activity of the purified expressed product was analyzed by indirect ELISA. The results indicated that enzyme digestion and DNA sequencing proved the prokaryotic vector to have been successfully constructed. SDS-PAGE analysis showed that the expressed protein was 27. 5 ku, and indirect ELISA demonstrated the purified protein had good immunological activity. In conclusion, the chloramphenicol ScFv with immunological activity was successfully expressed in prokaryotic cells, which laid the foundation for the establishment of drug residue detection methods for chloramphenicol.