Abstract
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.
Original language | English |
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Title of host publication | Methods in Molecular Biology |
Publisher | Humana Press Inc. |
Pages | 341-362 |
Number of pages | 22 |
DOIs | |
Publication status | Published - 2022 |
Publication series
Name | Methods in Molecular Biology |
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Volume | 2414 |
ISSN (Print) | 1064-3745 |
ISSN (Electronic) | 1940-6029 |
Bibliographical note
Publisher Copyright:© 2022, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
Keywords
- Complement system
- FPLC
- Human plasma
- IgG depletion
- IgM depletion