Purification, quantification, and functional analysis of complement factor H

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

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

5 Citations (Scopus)

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.

Original languageEnglish
Title of host publicationThe Complement System
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages207-223
Number of pages17
ISBN (Print)9781627037235
DOIs
Publication statusPublished - 2014
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1100
ISSN (Print)1064-3745

Keywords

  • Affinity chromatography
  • Complement
  • ELISA
  • Factor H
  • Factor I
  • Gel filtration
  • Protein G
  • Proteolysis
  • SDS-PAGE

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