食品科学 ›› 2009, Vol. 30 ›› Issue (11): 163-165.doi: 10.7506/spkx1002-6630-200911036

• 生物工程 • 上一篇    下一篇

氯霉素全抗原的合成与鉴定

董 斌,袁 茵,段 涛,林月霞,田素娟*   

  1. 广东药学院生命科学与生物制药学院
  • 收稿日期:2008-10-10 修回日期:2009-01-14 出版日期:2009-06-01 发布日期:2010-12-29
  • 通讯作者: 田素娟 E-mail:sjtian2005@yahoo.com.cn
  • 基金资助:

    广东省科技攻关计划项目(2006B20501005);广东省医学科学技术研究基金项目(A2006323);
    广州市科技计划项目(2006JI-C0221)

Synthesis and Identification of Complete Antigen of Chloramphenicol

DONG Bin,YUAN Yin,DUAN Tao,LIN Yue-xia,TIAN Su-juan*   

  1. School of Life and Biopharmacology, Guangdong Pharmaceutical University, Guangzhou 510006, China
  • Received:2008-10-10 Revised:2009-01-14 Online:2009-06-01 Published:2010-12-29
  • Contact: TIAN Su-juan E-mail:sjtian2005@yahoo.com.cn

摘要:

目的:为获得较高偶联率氯霉素全抗原,探索和优化碳二亚胺法(EDC)合成氯霉素(CAP)与牛血清白蛋白(BSA)全抗原合成条件。方法:采用单因素设计,优化影响合成的主要因素:蛋白浓度、CAP 与BSA 的摩尔比例、反应体系pH 值,通过紫外法和三硝基苯磺酸(TNBS)法测定合成后的偶联率。 结果:EDC 法合成氯霉素抗原较优条件为BSA 浓度15mg/ml、CAP:BSA=70:1、pH7.4,可得到偶联率为15 左右的氯霉素全抗原。结论:EDC法可用于制备较高偶联率的氯霉素全抗原,以用来进行抗体制备。

关键词: 氯霉素, 牛血清白蛋白, 碳二亚胺法, 偶联

Abstract:

Objective: To prepare a complete antigen of chloramphenicol (CAP) with high coupling rate, and optimize the coupling conditions of CAP and bovine serum albumin (BSA) by carbodiimide method. Method: Single-factor tests were designed to optimize three main factors affecting the synthesis such as protein concentration, mole ratio of CAP to BSA and pH in reaction system, and ultraviolet (UV) spectroscopy and TNBS method were employed to determine the coupling rate after the synthesis. Results: The optimum BSA concentration, mole ratio of CAP to BSA and pH were 15 mg/ml, 70:1 and 7.4, respectively. Under these conditions, the coupling rate of CPA antigen was about 15. Conclusion: The complete CPA antigen with high coupling rate can be prepared by carbodiimide method for the preparation of antibody.

Key words: chloramphenicol (CPA), bovine serum albumin (BSA), carbodiimide method, coupling

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