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图2 间接ELISA步骤 源自RayBiotech官网 夹心ELISA： 将捕获抗原的抗体（捕获抗体）附着在固相载体上，洗掉多余的未结合抗体后，添加抗原并被特异性捕获，通过直接ELISA或间接ELISA的形式检测抗原。这种组合进一步提高了检测的灵敏度和特异性，是最常见的ELISA实验形式。 一、ELISA技术概述 酶联免疫吸附测定（ELISA）是一种基于抗原抗体特异性反应的固相免疫检测技术，广泛应用于体液中微量物质的检测。它起源于20世纪70年代初，由多位学者共同推动发展，如今已成为分析化学和生物医学研究中的重要工具。 二、ELISA的核心原理 ELISA的基本原理是利用抗原与抗体之间. 间接 ELISA 间接 ELISA 类似于直接 ELISA；但主要的差别在于会通过第二偶联抗体来检测结合抗体。实验步骤的第一步便是将抗原固定在微孔板上。第二步是添加一种封闭试剂，防止结合抗体出现非特异性结合。第三，添加一种抗原特异性结合抗体，短暂孵育后，用缓冲液洗涤板孔，以去除多余的试剂.
抗原被动附着在固相载体（一般为96孔板）上，然后添加与酶（最常见的为HRP或AP）偶联的酶标一抗来检测抗原。这种检测方通常使用酶标仪来测定各样品孔吸光度，互相比较样品，或用已知分析物浓度制成的标准曲线来确定样… 酶联免疫吸附试验 (以下简称ELISA) ：是酶免疫测定技术中应用最广的技术。其基本方法是将已知的抗原或抗体吸附在固相载体 ( 聚苯乙烯微量反应板 ) 表面，使酶标记的抗原抗体反应在固相表面进行，用洗涤法将液相中的游离成分洗除。常用的 ELISA 法有双抗体夹心法和间接法，前者用于检测大分子. 一、ELISA技术原理及分类 酶联免疫吸附试验 （Enzyme-Linked Immunosorbent Assay, ELISA）是一种基于 抗原-抗体特异性结合 的高灵敏度免疫学检测技术。 其核心原理是将抗原或抗体的免疫学活性与酶催化底物显色的高效性相结合，通过颜色反应的强弱判断待测物浓度。 1.
受限于ELISA 灵敏度的限制, 在一些亲和力较弱(>100nM )的抗体抗原中,采用ELISA方法基本得不到有效的平台期,也无法测得准确的亲和力数值。
ELISA（Enzyme-Linked Immunosorbent Assay，酶联免疫吸附试验）是一种用于检测和定量蛋白质、抗体、抗原的免疫学方法，广泛应用于生物医学研究和诊断中。以下是详细的ELISA实验步骤、数据处理及作图过程的介绍： …
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