改变半抗原与载体蛋白的连接桥对提高水杨酸ELISA灵敏度的影响  

Effect of changing conjugation bridge between hapten and carrier protein on the sensitivity of salicylate elisa

作　　者：王树才[1] 周燮[1] 

机构地区：[1]南京农业大学农学系

出　　处：《中国免疫学杂志》1999年第1期37-39,共3页Chinese Journal of Immunology

摘　　要：目的：探讨水杨酸（ＳＡ）与载体蛋白间连接桥的改变对识别ＳＡ抗体的产生及ＳＡＥＬＩＳＡ灵敏度的影响。方法：将ＳＡ、４氨基水杨酸、５氨基水杨酸分别用不同的方法与载体蛋白相偶联，从而在ＳＡ苯环的Ｃ４、Ｃ５两个不同位点合成了３种免疫原：ＳＡ（Ｃ５）Ｎ＝ＮｐＡＨＡＢＳＡ（Ａ）、ＳＡ（Ｃ４）Ｎ＝ＣＨ（ＣＨ２）３ＣＨ＝ＮＢＳＡ（Ｂ）和ＳＡ（Ｃ５）ＮＨＣＨ２ＮＨＢＳＡ（Ｃ）及４种包被物：ＳＡ（Ｃ５）Ｎ＝ＮｐＡＨＡＯＶＡ、ＳＡ（Ｃ４）Ｎ＝ＣＨ（ＣＨ２）３ＣＨ＝ＮＯＶＡ、ＳＡ（Ｃ５）Ｎ＝ＣＨ（ＣＨ２）３ＣＨ＝ＮＯＶＡ、ＳＡ（Ｃ５）ＮＨＣＨ２ＮＨＯＶＡ。免疫新西兰白兔制备抗体，用ＥＬＩＳＡ法检测。结果：获得针对Ａ和Ｃ的两种多克隆抗体，后一种能专一识别游离态水杨酸，改变包被物中连接桥的结构可提高ＳＡＥＬＩＳＡ的检测灵敏度，故选用ＳＡ（Ｃ５）Ｎ＝ＣＨ（ＣＨ２）３ＣＨ＝ＮＯＶＡ建立了ＳＡＥＬＩＳＡ。结论：ＳＡ与载体蛋白间连接桥的改变对识别ＳＡ抗体的产生和ＳＡＥＬＩＳＡ的灵敏度有很大影响。Objective:To study the effect of changing conjugation bridge between hapten and carrier protein on the raising of antibodies against salicylic acid and the sensitivity of salicylate ELISA.Methods:Three kinds of salicylic acid (SA) immunogens:SA(C 5) N=N p AHA BSA(A)、SA(C 4) N=CH (CH 2) 3 CH=N BSA(B) and SA(C 5) NH CH 2 NH BSA(C) and four kinds of SA coating conjugates:SA(C 5) N=N p AHA OVA、SA(C 4) N=CH (CH 2) 3 CH=N OVA、SA(C 5) N=CH (CH 2) 3 CH=N OVA and SA(C 5) NH CH 2 NH OVA were synthesized by linking SA、4 aminosalicylic acid and 5 aminosalicylic acid respectively to bovine serum albumin(BSA) and ovalbumin(OVA) through different conjugation methods.Raising of antibodies in rabbits and effect of changing conjugation bridge on the sensitivity of saliaylate ELISA were detected by ELISA.Results:Two kinds of polyclonal antibodies were raised in rabbits against immunogens A and C respectively.The antibodies against immunogen C possessed a high specificity .Using these antibodies and choosing SA(C 5) N=CH (CH 2) 3 CH=N OVA as a coating conjugate,a kind of ELISA was developed which displayed a linearity range from 0.064 to 200 000 nmol of SA.Conclusion:Conjngation bridge changing has a great effect on the raising of antibodies against salicylic acid and the sensitivity of salicylate ELISA.

关 键 词：植物 水杨酸 半抗原 连接桥 ELISA 载体蛋白 

分 类 号：Q946.828.3[生物学—植物学] R446.61[医药卫生—诊断学]