婴幼儿配方乳粉中IgG活性质量浓度的测定  

Development of ELISA kit for detecting the bioactive content of IgG in infant formula milk powder

作　　者：侯方妮[1] 杜彦山 张志国 张佳程[1] 生庆海[3] 

机构地区：[1]青岛农业大学食品学院,山东青岛266109 [2]河北省乳业工程技术研究中心,石家庄050071 [3]东北农业大学国家乳业工业技术研究中心．哈尔滨150086

出　　处：《中国乳品工业》2009年第1期54-57,共4页China Dairy Industry

摘　　要：为经济、快速、准确的测定婴儿配方乳粉中IgG的活性质量浓度,建立了ELISA试剂盒。以质量浓度为5mg/L的兔抗牛IgG(37℃,3h)加4℃过夜包被酶标板,1︰250稀释度的HRP-兔抗牛IgG作为酶标二抗37℃温育1h,TMB底物溶液(150μL的TMB使用液+30μL的H2O2+10mL底物缓冲液)反应15min,酶联检测仪读出OD450nm值。应用上述条件制作试剂盒。结果表明,应用试剂盒制作的标准曲线方程为y=225.45x2-21.237x-1.4713,相关系数R2=0.9997,在4℃条件下保存至少2个月,回收率在0.93～1.06之间。应用试剂盒检测婴幼儿配方乳粉中的IgG,具有特异性强、灵敏、稳定和易于推广等优点。An economical, quick and exact ELISA kit was developed for determinadon of IgG in infant formula milk powder.Plates were coated with rabbit anti-bovine IgG 3 hours at 37 ℃ and overnight at 4 ℃.Dilution of enzyme-conjugated second antibody was 1 ： 250 and incubateted 1 hour at 37 ℃. Substrate/chromogen solution was prepared by adding 10 mL phophate-citrate buffer, 150 μL TMB soulution and 30 μL H2O2 and incubated 20 minutes.After stopping,plates were read at 450 run on a Titertek Multiskan MCC plate reader .Making use of aboveomentioned congditions produce ELISA kit.The calibration curve of bovine IgG by ELISA kit is γ = 225.45x^2 - 21.237x - 1.4713, R^2 is 0.9997.The ELISA kit can conserve an least 2 mongths at 4 ℃ and the average recovery is 0.93-1.06. The ELISA kit has following ad- vantages： highly specific, sensitive, stability and easy to extend.

关 键 词：IGG 婴幼儿配方乳粉 酶联免疫试剂盒 活性质量浓度 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]