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Purification, Quantification, and Functional Analysis of Complement Factor H

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The Complement System

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1100))

Abstract

Complement Factor H (FH) is an abundant, non-enzymic plasma/serum glycoprotein, which has a major role in regulating activation of the complement system. It can be purified from human plasma/serum by affinity chromatography, using a monoclonal anti-FH antibody as ligand. Other affinity chromatography ligands, including cardiolipin and trinitrophenyl-bovine serum albumin (TNP-BSA), can be used to purify human FH and also FH from a wide range of vertebrates, including mammals, birds, bony fish. Human FH protein concentration can be quantified by sandwich ELISA. The activity of FH is generally measured by assays which detect the cleavage, by complement factor I, of the complement protein C3b to form iC3b. Cleavage occurs only in the presence of a cofactor, and FH is one of a small number of cofactors for this reaction.

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Authors and Affiliations

  1. Department of Biochemistry, University of Oxford, Oxford, UK

    Bing-Bin Yu, Beryl E. Moffatt, Marina Fedorova, Claire G. S. Villiers, James N. Arnold, Eugenie Du, Astrid Swinkels, Man Chung Li & Robert B. Sim

  2. Department of Pharmacology, University of Oxford, Oxford, UK

    Ali Ryan

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  1. Bing-Bin Yu
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  2. Beryl E. Moffatt
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  3. Marina Fedorova
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  4. Claire G. S. Villiers
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  5. James N. Arnold
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  6. Eugenie Du
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  7. Astrid Swinkels
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  8. Man Chung Li
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  9. Ali Ryan
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  10. Robert B. Sim
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Editors and Affiliations

  1. Division of Infectious Diseases, Brigham and Women’s Hospital, Harvard University Harvard Medical School, Boston, Massachusetts, USA

    Mihaela Gadjeva

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© 2014 Springer Science+Business Media, New York

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Yu, BB. et al. (2014). Purification, Quantification, and Functional Analysis of Complement Factor H. In: Gadjeva, M. (eds) The Complement System. Methods in Molecular Biology, vol 1100. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-724-2_17

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  • DOI: https://doi.org/10.1007/978-1-62703-724-2_17

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-723-5

  • Online ISBN: 978-1-62703-724-2

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