Rapid Novor, Kitchener, ON, Canada.
Nat Commun. 2024 Oct 10;15(1):8790. doi: 10.1038/s41467-024-53105-8.
The antibody response to vaccination and infection is a key component of the immune response to pathogens. Sequencing of peripheral B cells may not represent the complete B cell receptor repertoire. Here we present a method for sequencing human plasma-derived polyclonal IgG using a combination of mass spectrometry and B-cell sequencing. We investigate the IgG response to the Moderna Spikevax COVID-19 vaccine. From the sequencing data of the natural polyclonal response to vaccination, we generate 12 recombinant antibodies. Six derived recombinant antibodies, including four generated with de novo protein sequencing, exhibit similar or higher binding affinities than the original natural polyclonal antibody. Neutralization tests reveal that the six antibodies possess neutralizing capabilities against the target antigen. This research provides insights into sequencing polyclonal IgG antibodies and the potential of our approach in generating recombinant antibodies with robust binding affinity and neutralization capabilities. Directly examining the circulating IgG pool is crucial due to potential misrepresentations by B-cell analysis alone.
接种疫苗和感染后的抗体反应是针对病原体的免疫反应的一个关键组成部分。外周 B 细胞的测序可能无法代表完整的 B 细胞受体库。在这里，我们提出了一种使用质谱和 B 细胞测序相结合的方法来对人血浆来源的多克隆 IgG 进行测序。我们研究了对 Moderna Spikevax COVID-19 疫苗的 IgG 反应。从接种疫苗的自然多克隆反应的测序数据中，我们生成了 12 种重组抗体。其中 6 种衍生的重组抗体，包括用从头蛋白测序生成的 4 种，表现出与原始天然多克隆抗体相似或更高的结合亲和力。中和试验表明，这 6 种抗体对靶抗原具有中和能力。这项研究为测序多克隆 IgG 抗体提供了新的见解，并且我们的方法在生成具有强结合亲和力和中和能力的重组抗体方面具有潜力。由于单独的 B 细胞分析可能存在代表性不足，因此直接检查循环 IgG 池至关重要。
