The ability to separate, identify, and quantify proteins from complex mixtures are key foundational methods across biochemistry teaching and research. In particular, enzyme‐linked immunosorbent assay (ELISA) is an important technique that is used to measure antigen concentrations in both industry and academia. There are four categories of ELISA, direct, indirect, competitive, and sandwich, each with their own applications. Sandwich ELISAs are used to determine antigen concentrations from complex mixtures of protein, such as a cell lysates, and are regularly used as medical diagnostics to diagnose illness and diseases ranging from hepatitis to celiac disease. One major problem with teaching the sandwich ELISA technique to students is the prohibitive cost due to the need to coat a 96‐well plate with a capture antibody. One solution to this problem would be to significantly reduce the role of each student in the lab, but this does not adequately prepare students to perform the procedure in a research or industry lab. Instead, this laboratory exercise teaches students the procedural knowledge needed to perform a direct sandwich ELISA, but uses a simulated experience performed within a wet‐lab environment. The presented scenario is the analysis of phosphorylated proteins within a synthetic signaling pathway, but because the lab uses simulated samples, it can be tailored to different topics and educational aims. The procedure is 10‐ to 26‐fold less expensive per student to deploy than an authentic sandwich ELISA. Students in the course report that the ELISA lab significantly strengthened the connection between theory and practice. © 2019 International Union of Biochemistry and Molecular Biology, 48(1):67–73, 2020.

中文翻译：

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用于对天然和工程细胞信号通路进行经济有效研究的模拟夹心酶联免疫吸附试验

从复杂混合物中分离、识别和量化蛋白质的能力是生物化学教学和研究的关键基础方法。特别是酶联免疫吸附测定 (ELISA) 是一种重要的技术，用于测量工业和学术界的抗原浓度。ELISA 有四类，直接、间接、竞争和夹心，每类都有自己的应用。夹心 ELISA 用于从复杂的蛋白质混合物（如细胞裂解物）中确定抗原浓度，并经常用作医学诊断，以诊断从肝炎到乳糜泻的疾病和疾病。向学生教授夹心 ELISA 技术的一个主要问题是成本过高，因为需要用捕获抗体包被 96 孔板。这个问题的一个解决方案是显着减少每个学生在实验室中的角色，但这并不能让学生为在研究或工业实验室中执行该程序做好充分的准备。相反，该实验室练习教授学生进行直接夹心 ELISA 所需的程序知识，但使用在湿实验室环境中进行的模拟体验。呈现的场景是分析合成信号通路中的磷酸化蛋白质，但由于实验室使用模拟样本，因此可以针对不同的主题和教育目标进行定制。与真正的夹心 ELISA 相比，每位学生部署该程序的成本要低 10 到 26 倍。课程中的学生报告说，ELISA 实验室显着加强了理论与实践之间的联系。