摘要
β-酪啡肽-7(β-Casomorphin-7)是一个有代表性的乳源外啡肽,有调节动物消化道运动和采食、内分泌、免疫等多种生物学功能。本研究以β-Casomorphin-7与人血清白蛋白(HSA)经戊二醛交联的复合物作为包被抗原;一抗为鸡抗β-Casomorphin-7与牛血清白蛋白(BSA)交联复合物的抗血清;酶标二抗为兔抗鸡IgG经戊二醛与辣根过氧化物酶(HRP)交联的复合物。包被抗原和一抗的稀释度经方阵稀释实验确定。本实验所建立的竞争ELISA方法的孔间差异为3.3%,板间差异为8.6%,检测灵敏度20ng/ml,检测范围20～320ng/ml,在实验浓度范围内。β-Casomor-phin-7的抗血清与β-内啡肽、脑啡肽、强啡肽、精氨酸降压素和神经降压素等活性肽未显示免疫交叉反应。表明所建立的方法有较高的特异性,可用于酪啡肽的体内功能研究。
Casomorphin -7,as one of representative exorphins derivedfrommilk,functioned physiologically on regulating food intake and gastroin testinal mobility,endocrine and im munity of animal.We tried to establi sh ELISA detection method forβ-Casomorphin -7.Nunc96-well microtiter plate was used as so lid -phase.The coating antigen on solid -phase wasβ-Casomorphin -7/HSA conjugation complex.The conce ntration for coating antigen and antibody were determined by dilu tion experiment.The result showed t hat detection forβ-Casomorphin -7with competitive inhibition ELISArepea table with lowbackground OD,inter well variation was about 3.3%,inter plate variation about 8.6%,detection sensitivity20ng /ml and the detectionrange20-320ng /ml.There was no immuno -across reactionofβ-Casomorphin -7,antibodies withβ-EP,EK,Dyn,AVPand NP.The results showedth at the methodwas of highspecificity and co uldbe used for studying onβ-Casomorphin -7s function.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2002年第2期94-97,共4页
Food Science
基金
国家自然基金资助(编号:30070565)
