基于聚丙烯酰胺凝胶电泳结合在线荧光成像的蛋白质特异性定量分析方法

蛋白质的特异性检测在生物分析和临床诊断中具有重要意义。现有方法如免疫固定电泳和蛋白质印迹法，需要手工染色和脱色等步骤，操作繁琐，耗时久且定量能力有限。本文结合在线荧光成像技术开发了一种蛋白质免疫聚丙烯酰胺凝胶电泳（PAGE）定量检测方法。该方法首先采用荧光标记抗体结合目标蛋白质，使用甲醛交联免疫复合物后进行电泳和在线成像，最终通过计算凝胶电泳图像中游离抗体的荧光信号强度实现定量分析。整个检测流程可在1.5 h内完成。本文以人转铁蛋白（transferrin，TRF）为模式蛋白进行方法学验证，结果显示该方法的线性范围为5.0~200.0 mg/L，线性相关系数（R2）为0.993 0，制作标准曲线的标准溶液中荧光信号强度的最大相对标准偏差（RSD）为1.65%，检出限为0.5 mg/L，加标回收率为98.2%~105.0%，表明该检测方法具有良好的精密度、灵敏度和准确度。相较于传统的PAGE方法，该方法利用实时荧光成像技术实现了对目标蛋白质的在线定量检测，具有分辨率高、特异性强、操作简便、分析快速以及重现性好等优点。该方法具有普适性，可应用于其他蛋白质的定量检测，在药物制备和临床诊疗等领域具有重要的应用价值。

• Publication year

  2025

• Venue

  Se pu = Chinese journal of chromatography

• Publication date

  2025-09-01

• Fields of study

  Not labeled

• Identifiers
  DOI 10.3724/SP.J.1123.2024.12017PMID 40910314PMCID 12412021
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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