Preparation of Chloramphenicol artificial immunogen and monoclonal antibodies

被引：0
作者：
Jian-Shu C. [1 ]
Hu-Ming Y. [1 ]
Yu Y. [1 ]
Min L. [1 ]
Guo-Qing Y. [1 ]
机构：
[1] Zhejiang University of Technology, College of Pharmaceutical Science, Hangzhou
来源：
Biotechnology and Biotechnological Equipment | 2011年 / 25卷 / 01期
关键词：
Artificial immunogen; Chloramphenicol; Monoclonal antibodies; Preparation;
D O I：
10.5504/bbeq.2011.0011
中图分类号：
学科分类号：
摘要：
Chloramphenicol (CAP) was coupled with bovine serum albumin (BSA) to prepare CAP artificial immunogen by mixed anhydride (MA) reaction, diazotization reaction and carbodiimide reaction whose reaction conditions were then further optimized. The successful linkage of the artificial immunogen was identified by UV Spectrophotometry (UV) and SDS Polyacrylamide gel electrophoresis (SDS-PAGE). Mice were immunized with different CAP artificial immunogen to obtain serum. Results determined by enzyme-linked immunosorbent assay (ELISA) showed that the CAP artificial immunogen synthesized by diazotization had the highest serum titer. Hybridomas were obtained by fusing mouse myeloma cells SP2/0 with splenocytes from the immunized mice. Hybridomas 2D1 secreting antibodies against CAP were obtained and subcloned. Ascites of monoclonal antibodies (McAb) were prepared by intraperitoneal injection of 1×106 cells of hybridomas 2D1 into mice abdomen. McAb obtained from hybridomas 2D1 were highly specific because there was no cross-reactivity with the drugs which were similar to CAP in their molecular structure.
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页码：2284 / 2289
页数：5
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