Tricine-SDS-PAGE电泳定量检测人体泪液中溶菌酶含量

史瑾，杨小琳，赵金礼*

（陕西慧康生物科技有限责任公司，陕西 西安，710054）

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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 kDa 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10～30 μg 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/mL。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。

关键词：人体泪液；溶菌酶；Tricine-SDS-PAGE 电泳；定量检测

中图分类号：R777.34 文献标志码：A文章编号：2096-1413(2018)02-0004-03

Quantitative detection of lysozyme in human tears by Tricine-SDS-PAGE electrophoresis
SHI Jin, YANG Xiao-lin, ZHAO Jin-li *
(Shaanxi Huikang Bio-tech Co., Ltd., Xi``an 710054, China)

ABSTRACT: Objective To establish a method for quantitative detection of lysozyme in human tears. Methods The protein distribution with the molecular weight between 3-30 kDa in human tear was analyzed by modified Tricine-SDS-PAGE electrophoresis to separate the tear lysozyme from other tear proteins. Gray scale analysis of lysozyme electrophoresis band was carried out by Image J software, which could quantitatively determinate the lysozyme. Results The tear lysozyme and other small molecule proteins could be separated effectively with modified Tricine -SDS -PAGE electrophores, the specificity was good for the detection of lysozyme in human tears. The sample size was within 10-30 μg, and the linear relationship between lysozyme content and electrophoretic, the specificity gray band was good (r=0.9934). There are 17 human 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 be applied for scientific research and clinical for the detection of lysozyme content in human tear.
KEYWORDS: human tears; lysozyme; Tricine-SDS-PAGE electrophoresis; quantitative detection

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