西北农业学报2025，Vol.34Issue(1)：133-139,7.DOI:10.7606/j.issn.1004-1389.2025.01.015
抗营养因子大豆凝集素夹心ELISA检测方法的建立
Establishment of a Sandwich ELISA for Detecting Soybean Agglutinin as an Anti-nutritional Factor
摘要
Abstract
To establish a sensitive and specific enzyme-linked immunosorbent assay(ELISA)for the detection of soybean agglutinin(SBA).Mouse-derived monoclonal antibody was used as the capturing antibody and rabbit-derived polyclonal antibody as the detecting antibody,with a sheep anti-rabbit en-zymic conjugate antibody as the secondary antibody.This sandwich method for SBA detection was op-timized in terms of antibody concentration and reaction time.This method was applied to detect SBA levels in differenet soybean products,including soybean,soybean meal,extruded soybean and enzyme hydrolyzed soybean meal.The results showed that the detection range of the developed sandwich ELISA was 39.06-1 250 ng/mL,the detection limit was 3.00 ng/mL,the fortified recovery was 77.96％-116.15％,the coefficient of variation was 6.11％-18.34％.There was no cross-reaction with other anti-nutritional factors such as trypsin inhibitors BBI,KTI,glycinin and β-conglycinin in soybean.The concentration of SBA in soybean was higher than that in soybean meal,extruded soy-bean meal and enzyme hydrolyzed soybean meal,respectively(P＜0.05).SBA in soybean meal was higher than in extruded soybean meal and enzyme hydrolyzed soybean meal,respectively(P＜0.05)and in extruded soybean meal was higher than in enzyme hydrolyzed soybean meal(P＜0.05).These findings indicate that the established SBA sandwich ELISA detection method is both sensitive and spe-cific and can be used for monitoring and detecting soybean agglutinin in feed.关键词
大豆凝集素/抗营养因子/多克隆抗体/单克隆抗体/夹心ELISAKey words
Soybean agglutinin/Anti-nutritional factor/Polyclonal antibody/Monoclonal antibody/Sandwich ELISA引用本文复制引用
胡骁飞,杨继飞,邢云瑞,庞杏豪,孙亚宁,魏凤仙..抗营养因子大豆凝集素夹心ELISA检测方法的建立[J].西北农业学报,2025,34(1):133-139,7.基金项目
国家重点研发计划(2018YFC1602903) （2018YFC1602903）
河南省现代农业产业技术体系项目(HARS-SS-12-S). The National Key Research and Development Program(No.2018YFC1602903) （HARS-SS-12-S）
Modern Agriculture Industry Technology System of Henan Province(No.HARS-SS-12-S). （No.HARS-SS-12-S）
