Division of Molecular Microbiology, School of Life Sciences, University of Dundee, Dundee, UK.
Methods Mol Biol. 2022;2414:37-45. doi: 10.1007/978-1-0716-1900-1_3.
Immunoprecipitation is an affinity purification technique that exploits the highly specific interactions formed between antibodies and their cognate antigens to purify molecules of interest from complex biological solutions. The generation of an effective humoral response provides protection against a wide range of gram-positive pathogens, and thus immunoprecipitation using antibodies purified from immune humans or animals provides a simple but effective means of isolating prospective vaccine antigens from fractionated bacterial cells for downstream identification. The commercial availability of antibody preparations from donated human plasma, containing antibodies against many common gram-positive pathogens, allows the protocol to be performed in the absence of bespoke vaccination experiments. Thus, immunoprecipitation has the potential to reduce the number of animals used in vaccine studies by allowing an initial screen for promising antigens to be conducted in vitro.
免疫沉淀是一种亲和纯化技术,利用抗体与其同源抗原之间形成的高度特异性相互作用,从复杂的生物溶液中纯化感兴趣的分子。有效的体液免疫反应为对抗广泛的革兰氏阳性病原体提供了保护,因此,使用从免疫的人类或动物中纯化的抗体进行免疫沉淀,为从分离的细菌细胞中分离潜在的疫苗抗原提供了一种简单而有效的方法,用于下游鉴定。来自捐赠人血浆的抗体制剂的商业可用性,含有针对许多常见革兰氏阳性病原体的抗体,使得该方案可以在没有定制疫苗实验的情况下进行。因此,免疫沉淀有可能通过允许在体外进行有希望的抗原的初步筛选,从而减少疫苗研究中使用的动物数量。