Purification of Human Complement Component C4 and Sample Preparation for Structural Biology Applications

Alessandra Zarantonello, Sofia Mortensen, Nick S. Laursen, Gregers R. Andersen*

*Corresponding author for this work

Research output: Contribution to book/anthology/report/proceedingBook chapterResearchpeer-review

2 Citations (Scopus)

Abstract

Activated complement component C4 (C4b) is the nonenzymatic component of the classical pathway (CP) convertases of the complement system. Preparation of C4 and C4b samples suitable for structural biology studies is challenging due to low yields and complexity of recombinant C4 production protocols reported so far and heterogeneity of C4 in native sources. Here we present a purification protocol for human C4 and describe sample preparation methods for structural investigation of C4 and its complexes by crystallography, small angle X-ray scattering, and electron microscopy.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
Volume2227
Place of publicationNew York
PublisherHumana Press
Publication date2021
Pages249-264
ISBN (Print)978-1-0716-1015-2
ISBN (Electronic)978-1-0716-1016-9
DOIs
Publication statusPublished - 2021
SeriesMethods in Molecular Biology
Volume2227
ISSN1064-3745

Keywords

  • Complement C4
  • Gel filtration
  • Ion exchange
  • Negative stain electron microscopy
  • Small angle X-ray scattering
  • X-ray crystallography

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