食品科学 ›› 2010, Vol. 31 ›› Issue (22): 450-452.doi: 10.7506/spkx1002-6630-201022101

• 分析检测 • 上一篇    下一篇

竞争ELISA 法对牛乳β - 酪蛋白的检测

宋宏新,潘 洁,柏红梅,薛海燕   

  1. 陕西科技大学生命科学与工程学院
  • 收稿日期:2010-07-21 出版日期:2010-11-25 发布日期:2010-12-29
  • 通讯作者: 宋宏新 E-mail:hongxinsong@163.com
  • 基金资助:

    陕西省科学技术厅农业攻关项目(2009K02-09);陕西省教育厅产业化培育项目(09JC19)

Detection of Bovine β-Casein by Competitive ELISA

SONG Hong-xin,PAN Jie,BAI Hong-mei,XUE Hai-yan   

  1. Institute of Life Science and Engineering, Shaanxi University of Science and Technology, Xi’an 710021, China
  • Received:2010-07-21 Online:2010-11-25 Published:2010-12-29
  • Contact: SONG Hong-xin E-mail:hongxinsong@163.com

摘要:

通过确定包被抗体最佳稀释度(anti- β-CN IgY 1:160)、封闭液及封闭时间(含1% 明胶的PBST,封闭1h)、酶标抗原(1:20)和待测脱脂牛乳样品稀释液(1:80)抗体作用时间(60min)、底物最佳反应时间(20min)、温度(37℃)等对竞争ELISA 效果有重要影响的因素,建立检测牛乳β- 酪蛋白的竞争ELISA 法。经敏感性实验和重复性实验证明本实验建立的竞争ELISA 法最低检出量为5.1μg/mL,变异系数(CV)小于5%,可以开发应用于牛乳及其制品的蛋白质品质快速检测。

关键词: 竞争ELISA, β- 酪蛋白, 鸡卵黄抗体(IgY)

Abstract:

In order to establish a competitive ELISA method for detecting β-casein of bovine milk, the factors to affect competitive-ELISA were investigated. The optimal conditions for competitive ELISA method were coated anti-β-CN IgY concentration of 1:160, 1% gelatin-PBST as the blocking solution and blocking time of 1 h. The working titers betweenβ-CNHRP and detection samples were 1:20 and 1:80, respectively. The reaction time of antibody and antigen was 1 h and the optimum enzymatic reaction time was 20 min. Detection results indicated that the detected limit was 3.5% with a variation coefficient (CV) of less than 5%. This method can be used for rapid and accurate detection ofβ-casein in bovine milk and dairy products.

Key words: competitive ELISA, β-casein, egg-yolk antibody

中图分类号: