抗体
免疫系统
琼脂糖
人血浆
免疫学
补体系统
亲和层析
病毒学
生物
化学
色谱法
生物化学
酶
作者
Frances Alexander,Emily Brunt,Holly E. Humphries,Breeze E. Cavell,Stephanie Leung,Lauren Allen,Rachel Halkerston,E. Lesné,Elizabeth J. Penn,Stephen R. Thomas,Andrew Gorringe,Stephen Taylor
出处
期刊:Methods in molecular biology
日期:2021-11-17
卷期号:: 341-362
被引量:7
标识
DOI:10.1007/978-1-0716-1900-1_18
摘要
Complement is a key component of functional immunological assays used to evaluate vaccine-mediated immunity to a range of bacterial and viral pathogens. However, standardization of these assays is complicated due to the availability of a human complement source that lacks existing antibodies acquired either through vaccination or natural circulation of the pathogen of interest. We have developed a method for depleting both IgG and IgM in 200 mL batches from pooled hirudin-derived human plasma by sequential affinity chromatography using a Protein G Sepharose column followed by POROS™ CaptureSelect™ IgM Affinity resin. The production of large IgG- and IgM-depleted batches of human plasma that retains total hemolytic and alternative pathway activities allows for improved assay standardization and comparison of immune responses in large clinical trials.
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