Milchram Lisa, Soldo Regina, Regele Valerie, Schönthaler Silvia, Degeorgi Markus, Baumgartner Sophie, Kopp Elena, Weinhäusel Andreas
Competence Unit Molecular Diagnostics, Center for Health and Bioresources, Austrian Institute of Technology, Giefinggasse 4, Vienna, 1210, Austria.
Biotechniques. 2022 Apr;72(4):134-142. doi: 10.2144/btn-2021-0107. Epub 2022 Mar 2.
ELISA is the current standard for (auto)antibody diagnostics. Once established, ELISA protocols can be easily adapted for novel antigens; however, peptide-based protocols are rarely available. Herein the authors describe the results of a technical investigation of an indirect ELISA protocol using peptides conjugated onto a protein carrier based on click chemistry and immobilized in standard plastics. The authors compared this approach with the common biotin-avidin system and obtained a slightly improved limit of detection for purified IgG of 25-100 ng/well compared with 25-1000 ng/well. Reproducibility and stability of the methodological approach were conducted for further technical characterization. Indirect ELISA using immunoreactive peptides conjugated to bovine serum albumin offers a reliable method that is complementary to standard plastics and plate readers.
酶联免疫吸附测定（ELISA）是目前（自身）抗体诊断的标准方法。一旦确立，ELISA方案可轻松适用于新型抗原；然而，基于肽的方案却很少见。在此，作者描述了一项技术研究的结果，该研究采用基于点击化学将肽缀合到蛋白质载体上并固定在标准塑料制品中的间接ELISA方案。作者将此方法与常见的生物素-抗生物素蛋白系统进行了比较，结果显示，对于纯化的IgG，检测限略有提高，从每孔25 - 1000纳克提高到每孔25 - 100纳克。为进行进一步的技术特性分析，对该方法的重现性和稳定性进行了研究。使用与牛血清白蛋白缀合的免疫反应性肽的间接ELISA提供了一种可靠的方法，可作为标准塑料制品和酶标仪的补充。
