Tricine-SDS-PAGE电泳定量检测人体泪液中溶菌酶含量  

Quantitative detection of lysozyme in human tears by Tricine-SDS-PAGE electrophoresis

作　　者：史瑾 杨小琳 赵金礼 

机构地区：[1]陕西慧康生物科技有限责任公司,陕西西安710054

出　　处：《临床医学研究与实践》2018年第2期4-6,共3页Clinical Research and Practice

摘　　要：目的建立一种定量检测人体泪液中溶菌酶含量的方法。方法通过改进的Tricine-SDS-PAGE电泳,分析正常人体天然泪液中分子量在3~30 k Da的蛋白质分布,使泪液溶菌酶与其它泪液蛋白有效分离,用Image J软件对溶菌酶电泳条带进行灰度分析,定量测定其中溶菌酶的含量。结果泪液溶菌酶和其它小分子蛋白可用改进的Tricine-SDS-PAGE电泳有效分离,对于人体泪液中的溶菌酶检测专属性良好;上样量10~30μg范围内,溶菌酶含量和电泳条带灰度面积线性关系良好(r=0.9934);分析17例人体泪液的样本,其中溶菌酶的含量在1.38~3.07 mg/m L。结论改进的Tricine-SDS-PAGE电泳法简单直观,并能批量性的一次分析多个样本,可用于科研及临床上人体泪液中溶菌酶的含量检测。Objective To establish a method for quantitative detection of lysozyme in human tears. Methods The proteindistribution with the molecular weight between 3-30 kDa in human tear was analyzed by modified Tricine-SDS-PAGEelectrophoresis to separate the tear lysozyme from other tear proteins. Gray scale analysis of lysozyme electrophoresis bandwas carried out by Image J software, which could quantitatively determinate the lysozyme. Results The tear lysozyme andother small molecule proteins could be separated effectively with modified Tricine -SDS -PAGE electrophores, thespecificity was good for the detection of lysozyme in human tears. The sample size was within 10-30 滋g, and the linearrelationship between lysozyme content and electrophoretic, the specificity gray band was good （r=0.9934）. There are 17human natural tear samples were analyzed, and the content of lysozyme was 1.38-3.07 mg/mL. Conclusion The Tricine-SDS-PAGE electrophoresis is simple and direct, and multiple samples could be analyzed in batch at one time. It can beapplied for scientific research and clinical for the detection of lysozyme content in human tear.

关 键 词：人体泪液 溶菌酶 Tricine-SDS-PAGE电泳 定量检测 

分 类 号：R777.34[医药卫生—眼科]