Tumor Glycomics Laboratory, SRI International Biosciences Division, Menlo Park, CA, USA.
Methods Mol Biol. 2023;2612:129-139. doi: 10.1007/978-1-0716-2903-1_10.
Carbohydrates have a great potential in generating structural and immunological diversities. Microbial pathogens often decorate their outmost surfaces with specific carbohydrate signatures. Carbohydrate antigens also differ significantly from protein antigens in physiochemical properties, especially in surface display of antigenic determinants in aqueous solutions. Technical optimization or modifications are often needed when we apply standard procedures for protein-based enzyme-linked immunosorbent assay (ELISA) to assess immunologically potent carbohydrates. We present here our laboratory protocols for performing carbohydrate ELISA and discuss several assay platforms that may be applied complementarily to explore the carbohydrate moieties that are critical for host immune recognition and induction of glycan-specific antibody responses.
碳水化合物在产生结构和免疫多样性方面具有巨大的潜力。微生物病原体经常在其最外层表面装饰上特定的碳水化合物特征。碳水化合物抗原在理化性质上也与蛋白质抗原有很大的不同，特别是在抗原决定簇在水溶液中的表面展示。当我们应用基于蛋白质的酶联免疫吸附测定（ELISA）的标准程序来评估具有免疫活性的碳水化合物时，通常需要进行技术优化或修改。我们在此介绍进行碳水化合物 ELISA 的实验室方案，并讨论几种可能互补应用的检测平台，以探索对宿主免疫识别和诱导糖特异性抗体反应至关重要的碳水化合物部分。
