麦胚凝集素红细胞凝集活性检测方法的改进.pdfVIP

第 23 卷第 1 期 无 锡 轻 工 大 学 学 报 Vol. 23 　No. 1 　 　 　 　　　　 　 2004 年 1 月 Journal of Wuxi University of Light Industry 　 　 　　　　 Jan. 　2004 ( ) 　文章编号 :1009038X 2004 麦胚凝集素红细胞凝集活性检测方法的改进 单 良 , 　姚惠源 (江南大学 食品学院 ,江苏 无锡 214036) 摘 　要 : 对红细胞进行蛋白酶处理、戊二醛固定 ,采用 p H 7. 2 等渗磷酸盐作为缓冲液 ,用光径 0. 5 cm 比色池测定凝集活性,读取测定终点的时间以 2 h 为宜. 建立的检测方法简单易行 ,得出的结果 具有良好的灵敏度及重现性. 关键词 : 麦胚凝集素 ; 红细胞凝集活性 ; 检测方法 中图分类号:R 446 文献标识码 : A An Improved Protocol for Determination of Hemagglutinating Activity of Wheat Germ Agglutinin SHAN Liang , 　YAO Huiyuan ( School of Food Science and Technology , Southern Yangtze University , Wuxi 214036 , China) Abstract : The procedure for determination of hemagglutination activity ( HA ) of wheat germ agglutinin ( W GA ) was improved. Major improvements include : 1 ) . the self sedimentation of erythrocytes was deducted in the definition and calculation of HA ; 2) . erythrocytes were sensitized with trypsin and fixed with glutaraldehyde prior to HA tests ; 3) . p H 7. 2 isotonic phosphate buffered saline was used as the working liquid ; 4) . UV absorption of erythrocyte suspensions was taken using 0. 5 cm pathlength cells ; and 5) . appropriate endpoint was determined as 2 hours after adding W GA into the suspension. The protocol thus derived was proved to be convenient , capable of having good sensitivity and reproducibilit y. Key words : wheat germ agglutinin (W GA) ; hemagglutinating activity ( HA) ; determination (