Generation of a Universal Human Complement Source by Large-Scale Depletion of IgG and IgM from Pooled Human Plasma

Frances Alexander; Emily Brunt; Holly Humphries; Breeze Cavell; Stephanie Leung; Lauren Allen; Rachel Halkerston; Elodie Lesne; Elizabeth Penn; Stephen Thomas; Andrew Gorringe; Stephen Taylor

doi:10.1007/978-1-0716-1900-1_18

Generation of a Universal Human Complement Source by Large-Scale Depletion of IgG and IgM from Pooled Human Plasma

Frances Alexander
, Emily Brunt
, Holly Humphries
, Breeze Cavell
, Stephanie Leung
, Lauren Allen
, Rachel Halkerston
, Elodie Lesne
, Elizabeth Penn
, Stephen Thomas
, Andrew Gorringe
, Stephen Taylor^*

^*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

11 Citations (Scopus)

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
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	341-362
Number of pages	22
DOIs	https://doi.org/10.1007/978-1-0716-1900-1_18
Publication status	Published - 2022

Publication series

Name	Methods in Molecular Biology
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.

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

Complement system
FPLC
Human plasma
IgG depletion
IgM depletion

Access to Document

10.1007/978-1-0716-1900-1_18

Cite this

Alexander, F., Brunt, E., Humphries, H., Cavell, B., Leung, S., Allen, L., Halkerston, R., Lesne, E., Penn, E., Thomas, S., Gorringe, A., & Taylor, S. (2022). Generation of a Universal Human Complement Source by Large-Scale Depletion of IgG and IgM from Pooled Human Plasma. In Methods in Molecular Biology (pp. 341-362). (Methods in Molecular Biology; Vol. 2414). Humana Press Inc.. https://doi.org/10.1007/978-1-0716-1900-1_18

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doi = "10.1007/978-1-0716-1900-1\_18",

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Alexander, F, Brunt, E, Humphries, H , Cavell, B , Leung, S , Allen, L, Halkerston, R, Lesne, E, Penn, E, Thomas, S , Gorringe, A & Taylor, S 2022, Generation of a Universal Human Complement Source by Large-Scale Depletion of IgG and IgM from Pooled Human Plasma. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 2414, Humana Press Inc., pp. 341-362. https://doi.org/10.1007/978-1-0716-1900-1_18

Generation of a Universal Human Complement Source by Large-Scale Depletion of IgG and IgM from Pooled Human Plasma. / Alexander, Frances; Brunt, Emily; Humphries, Holly et al.
Methods in Molecular Biology. Humana Press Inc., 2022. p. 341-362 (Methods in Molecular Biology; Vol. 2414).

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

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T1 - Generation of a Universal Human Complement Source by Large-Scale Depletion of IgG and IgM from Pooled Human Plasma

AU - Alexander, Frances

AU - Brunt, Emily

AU - Humphries, Holly

AU - Cavell, Breeze

AU - Leung, Stephanie

AU - Allen, Lauren

AU - Halkerston, Rachel

AU - Lesne, Elodie

AU - Penn, Elizabeth

AU - Thomas, Stephen

AU - Gorringe, Andrew

AU - Taylor, Stephen

PY - 2022

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AB - 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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KW - FPLC

KW - Human plasma

KW - IgG depletion

KW - IgM depletion

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T3 - Methods in Molecular Biology

SP - 341

EP - 362

BT - Methods in Molecular Biology

PB - Humana Press Inc.

ER -

Generation of a Universal Human Complement Source by Large-Scale Depletion of IgG and IgM from Pooled Human Plasma

Abstract

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Bibliographical note

UN SDGs

Keywords

Access to Document

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