安徽农业科学2016，Vol.44Issue(33)：74-76,102,4.
间接酶联免疫吸附法测定大鲵凝集素
An Indirect Non-compe titive Enzyme-linked Immunosorbent Asas y for Determin ation of Andrias davidianus Lectin
摘要
Abstract
Objective] To prepare polyclonal antibody against the content of lectin in skin mucous of Andrias davidianus and develop an indirect non-competitive enzyme-linked immunosorbent assay (ELISA).[Method] Andrias davidianus lectin was used as complete antigen.The New Zeal-and white rabbits were immunized according to the immunization procedure .Finally through the optimization of coating amount of lectin and anti-body dilution,an indirect non-competitive enzyme-linked immunosorbent assay was developed .[ Result] The titer of the antiserum determined by ELISA was up to 1∶16 000.Determined polyclonal antibody diluted was 1∶3 000.The best working dilution of horseradish peroxidase-conjugated affinipure goat anti rabbit IgG was 1∶30 000.The linear range of the method was 31.25-1 000.00 ng/mL (R2 >0.99).The variation coefficients of intra -assay and inter-assay were 2.50%-9.88% and 2.69%-11.59%,respectively.Recoveries ranged from 91.5% to 109.5%.[Conclu-sion] A rapid,sensitive and repeatable indirect non-competitive ELISA detection method for An drias davidianus lectin allergens was established successfully.This method is suitable for qualitative determination of Andrias davidianus lectin.关键词
大鲵/凝集素/酶联免疫/多克隆抗体Key words
Andrias davidianus/Lectin/Enzyme-linked immunosorbent assay/Polyclonal antibody分类
农业科技引用本文复制引用
王晓龙,佟长青,李伟..间接酶联免疫吸附法测定大鲵凝集素[J].安徽农业科学,2016,44(33):74-76,102,4.基金项目
国家自然基金项目（31571916）；大连海洋大学引进人才及在职培养博士启动项目（100914262）。 （）
