摘要
基于实验室开发的蛋白电泳分析仪,该文提出了一种可实现蛋白质分子SDS-PAGE分离,以及分子量和浓度在线分析的方法。通过对10.5~107 kDa蛋白质Marker进行电泳,并在线采集其凝胶电泳图像,分析其灰度值与蛋白质分子浓度间的关系。结果表明:该设备可在30 min内实现小于107 kDa的蛋白质样品的快速分离,当紫外光照射5 min时各蛋白条带荧光强度达到峰值;在10.5~107 kDa范围内,其迁移距离与分子量成正比,相关系数可达0.93;高于20.1 kDa的蛋白质分子,其荧光强度与质量浓度在0.72~3.96μg/μL范围内呈线性关系,相关系数均高于0.988;采用开发的蛋白电泳分析仪对泪液中的蛋白成分进行分析,并成功分离出免疫球蛋白A、乳铁蛋白、血清蛋白、脂质运载蛋白和溶菌酶。该研究对蛋白质分子快速检测设备的开发具有重要的应用价值。与传统的蛋白质凝胶电泳方法相比,开发的蛋白电泳分析仪可将蛋白质的电泳时间由1.5 h以上缩短至30 min内。该研究对于蛋白质分子快速检测设备的开发具有重要的应用价值。
Based on the self-built protein electrophoresis instrument,this paper proposes a method that can realize the SDS-PAGE separation of proteins,and online analysis of its molecular weight and concentration.Through electrophoresis of 10.5-107 kDa protein Marker,and the relationship between its gray value and protein molecular concentration is analyzed by analysis of its gel electrophoresis images.Results showed that:the device can achieve rapid separation of protein samples with a molecular weight of less than 107 kDa within 30 min,and the fluorescence intensity of each protein band reaches its peak when irradiated with ultraviolet light for 5 min;Within the range of 10.5-107 kDa,the migration distance is directly proportional to the molecular weight,with a correlation coefficient of up to 0.93;For protein molecules above 20.1 kDa,the fluorescence intensity shows a linear relationship in the concentration range of 0.72-3.96μg/μL with with correlation coefficients more than 0.988;Using the developed protein electrophoresis instrument analyzing the protein components in tears,and immunoglobulin A,lactoferrin,serum protein,lipid transport protein,and lysozyme were successfully separated from tears.Compared with the traditional protein gel electrophoresis methods,the developed self-built protein electrophoresis instrument could shorten the protein electrophoresis time from over 1.5 h to within 30 min.This study was important for the development of rapid protein molecule detection equipment.
作者
樊帅强
游庆祥
曾媛
杨波
马鸣远
张大伟
李振庆
FAN Shuai-qiang;YOU Qing-xiang;ZENG-yuan;YANG Bo;MA Ming-yuan;ZHANG Da-wei;LI Zhen-qing(School of Optical-Electrical and Computer Engineering,Key Laboratory of Medical Optical Instrument and Equipment of Ministry of Education,University of Shanghai for Science and Technology,Shanghai 200093;Changzhou Institute of Technology,Changzhou 5650871,China;Shanghai Key Laboratory of Molecular Imaging,Shanghai University of Medicine and Health Sciences,Shanghai 201318,China)
出处
《分析测试学报》
北大核心
2025年第10期2204-2208,共5页
Journal of Instrumental Analysis
基金
上海市“科技创新行动计划”生物医药科技支撑专项(19ZR1477500)。
