重组毕赤酵母AiiA蛋白表达量的双抗体夹心ELISA测定  

Development of a Sandwich Enzyme-linked Immunosorbent Assay for Quantitative Analysis on the Recombinant AiiA Expressed by Pichia pastoris

作　　者：曾世涌[1] 简思美 谢盼盼[1] 杨梅[1] 

机构地区：[1]福建师范大学生命科学学院,福建福州350117

出　　处：《福建师范大学学报（自然科学版）》2014年第6期56-61,共6页Journal of Fujian Normal University：Natural Science Edition

基　　金：福建省自然科学基金资助项目(2012J01123);福建省教育厅资助项目(JK2012009)

摘　　要：利用双抗体夹心ELISA的方法，建立了能对重组毕赤酵母表达AiiA蛋白进行定量测定的方法．双抗体夹心ELISA方法建立的标准AiiA蛋白曲线显示，随着AiiA蛋白质量浓度增大，显色后A410值增大，在AiiA质量浓度为1．11—35．5μg·mL-1的范围内具有良好的线性关系．经方法学分析表明，该方法对合有AiiA蛋白的发酵样品的检测具有良好的特畀性和精确度，AiiA的检测限为0．80μg·mL-1，批内差畀系数和批间差异系数分别为5．22％和10．30％．测定8．86，2．22，1．11μg·mL-13个质量浓度回收率分别为104％、97．3％和102％，能够满足定量要求．A simple, sensitive and effective quantitative method based on double-antibody sandwich enzyme-linked immunosorbent assay （ELISA） was established for estimating the concen- tration of AiiA expressed by Pichia pastoris in fermentation supernatant. The results showed that the absorbance at 410 nm increased progressively as the concentration of AiiA increased, which was lin- early correlated as the concentration of AliA increased from 1.11 μg mL- 1 to 35.5 μg mL- 1 The detectable limitation is 0. 80 μg - mL- 1 of AiiA protein, and the intra-assay and inter-assay co- efficient of variation were 5.22% and 10. 30% , respectively. The recoveries at three concentrations of 8.86, 2.22, 1.11 μg mL-1 were 104% , 97.3% and 102% respectively. The results showed that the double-antibody sandwich ELISA was sensitive and specific for quantitative analysis of AiiA protein.

关 键 词：酶联免疫吸附测定 AiiA蛋白 双抗体夹心法 

分 类 号：Q936[生物学—微生物学]