江苏大学学报（医学版）2015，Vol.25Issue(2)：98-102,5.DOI:10.13312/j.issn.1671-7783.y140079
多西环素多克隆抗体的制备及鉴定
Preparation and identification of doxycycline polyclonal antibody
摘要
Abstract
Objective:To conjugate the doxycycline (Dox) with bull serum albumin (BSA)/ovalbumin (OVA) and generate polyclonal antibody against the Dox.Methods:The Dox was coupled with BSA/OVA via direct,one-step formaldehyde,diazo plus mixed anhydride and diazo plus carbodiimide conjugation method,respectively.The BALB/c mice were immunized with Dox-BSA which conjugated by most efficient method.The titer,sensitivity and specificity of antibody were detected and assayed by direct or indirect competitive inhibition ELISA.Results:It had been declared that the diazo plus carbodiimide conjugation method was most efficient,and the coupling ratio of Dox ∶ BSA was 8.37 ∶ 1,and Dox ∶ OVA was 4.92 ∶ 1.The polyclone antibody (pAb) titer was more than 1 ∶ 8 000 by indirect ELISA.The half maximal inhibitory concentration (IC50) was 98.89-120.32 μg/L.There was less the cross reactivity with other tetracyclines.Conclusion:The conjugation method of diazo plus carbodiimide was the most efficient and the pAb has high titer,better sensitivity and specificity.关键词
多西环素/多克隆抗体/偶联/鉴定Key words
doxycycline/polyclonal antibody/coupling/identification分类
医药卫生引用本文复制引用
王军,尹蕾,蒋砚秋,禚娅,刘霞,王荟,张苗苗,杜凤移,夏圣..多西环素多克隆抗体的制备及鉴定[J].江苏大学学报（医学版）,2015,25(2):98-102,5.基金项目
国家自然科学基金资助项目(81273202) （81273202）
