摘要
分别以免血清IgG与鸡卵黄抗体（IgY）为检测抗体,建立了两种牛乳κ-酪蛋白的间接ELISA定量方法.牛乳κ-酪蛋白分别免疫新西兰大白兔和临产母鸡,经亲和层析分离纯化后得到anti-κ-CN-IgG与anti-κ-CN-IgY两种抗体.两种抗体均与β-酪蛋白、α-乳白蛋白和乳清粉存在一定交叉反应,抗体与相应抗原进行免疫吸收后得到高特异性IgG与IgY.用吸收后的anti-κ-CN-IgG建立的间接ELISA法,当标准κ-CN质量浓度在0.098～3.125 mg/L时,A450 nm值与κ-CN的质量浓度具有线性关系,变异系数小于1％,回收率在99.10％～101.1％之间；用吸收后的anti-κ-CN-IgY检测κ-CN,在0.098～6.25 mg/L时,A450 nm值与κ-CN的质量浓度成良好的线性关系,变异系数小于2％,回收率在98％～100.8％之间.当三聚氰胺掺假量在0～20％时,两种抗体均能通过κ-CN的定量检测而实现乳品的掺假检验.
Two indirect ELISA techniques were reported to detect bovine K--casein （K-CN） based on chicken IgY and rabbti serum IgG. Bovine K--casein was used as antigen to inject white rabbit and chicken. Rabbit anti-K-CN-IgG and chicken anti-K-CN-IgY were purified from rabbit sera and chicken egg yolk by affinity chromatography. This two antibodies are all have cross reactions with β-casein, α- lactoalbumin and whey powder and their specificity was improved by the Immune absorption technique. The two post-absorbed antibodies were used to quantify bovine K-CN in indirect ELISA. The method based on IgG present good linear relationship with A450nm value when the K-CN concentration ranged from 0.098 to 3.125 ug/mL with linear coefficient of 0.9805, CV less than 1% and recovery in range of 99.10%- 101.1%; while the method based on IgY present good linear relationship with A450nm value as well when the K-CN concentration ranged from 0.098 to 6.25 ug/mL with linear coefficient of 0.9901, CV less than 2% and recovery in range of 98.0%-100.8 %. The repeatabilities of two metheds are good enough to quantify K--CN in milk products. IgY are suitable to replace IgG using in quantity detection. This article provided a new technology to quantify K-CN. The two antibodies can used to determine the adulteration of milk through quantifying K-CN by the established indirect ELISA, when melamine adulteration quantity are ranged at 0-20%.
出处
《中国乳品工业》
CAS
北大核心
2014年第2期48-51,共4页
China Dairy Industry
基金
国家自然科学基金项目(31301405)
陕西省科学技术研究发展计划项目(2012JM2009)
陕西省教育厅自然科学基金专项项目(12JK0655)
