食品科学

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莱克多巴胺人工抗原的合成与鉴定

邓发亮,刘晓云,唐时幸,王继华   

  1. 广州万孚生物技术股份有限公司,广东 广州 510663
  • 出版日期:2013-09-25 发布日期:2013-09-27

Synthesis and Identification of Stable Artificial Ractopamine Antigen

DENG Fa-liang,LIU Xiao-yun,TANG Shi-xing,WANG Ji-hua   

  1. Guangzhou Wondfo Biotechnology Co. Ltd., Guangzhou 510663, China
  • Online:2013-09-25 Published:2013-09-27

摘要:

为合成新的、稳定的莱克多巴胺人工抗原,对莱克多巴胺结构进行修饰,合成带羧基基团的莱克多巴胺半抗原,经碳二亚胺法与载体BSA蛋白偶联制备莱克多巴胺人工抗原,采用紫外光谱扫描法、蛋白质电泳法和胶体金免疫层析法检测偶联物。莱克多巴胺半抗原的结构经气相质谱联用仪确证。结果表明:莱克多巴胺人工合成抗原连接成功,与抗莱克多巴胺抗体有特异性反应,具有抗原性。该方法合成的莱克多巴胺人工抗原结构未见报道、稳定性优越,可作为莱克多巴胺免疫检测方法中包被抗原用于莱克多巴胺检测,也可作为免疫源性抗原用于制备抗莱克多巴胺抗体。

关键词: 莱克多巴胺, 半抗原, 人工抗原, 免疫检测

Abstract:

A novel and stable artificial ractopamine hapten with the active group carboxyl group was designed and
synthesized by chemical modification, and linked to bovine serum albumin through ethyl carbodiimide hydrochloride
method to form artificial ractopamine antigen. The conjugation was confirmed by UV scanning method and protein
electrophoresis. The binding reaction of the artificial ractopamine antigen with anti-ractopamine antibody was verified by
gold immunochromatography assay. The ractopamine hapten was characterized by gas chromatography-mass spectrometry.
The ractopamine-BSA conjugation was stable and active for at least 6 months at 4 ℃. Our results indicate that ractopamine-
BSA can be successfully synthesized by the above method, and can be used as coating antigen in immunoassay for
ractopamine detection. In addition, we found that the synthesized antigen could be used as an immunogen to produce antiractopamine
antibody.

Key words: ractopamine, hapten, artificial antigen, ractopamine, immunoassay