Fast IMAC purification of non-tagged S100A8/A9 (calprotectin) from Homo sapiens and Sus scrofa

Protein Expr Purif. 2023 Aug:208-209:106275. doi: 10.1016/j.pep.2023.106275. Epub 2023 Apr 19.

Abstract

S100A8/A9 (calprotectin) is a damage-associated molecular pattern molecule (DAMP) that plays a key role in the innate immune response of mammalia. S100A8/A9 is therefore widely used as a biomarker in human and veterinary medicine, but diagnostic tools for the detection of S100A8/A9 are rarely optimised for the specific organism, since the corresponding S100A8/A9 is often not available. There is need for an easy, reliable protocol for the production of recombinant, highly pure S100A8/A9 from various mammalia. Here we describe the expression and purification of recombinant human and porcine S100A8/A9 by immobilized metal affinity chromatography (IMAC), which takes advantage of the intrinsic, high-affinity binding of native un-tagged S100A8/A9 to metal ions. Highly pure S100A8/A9 is obtained by a combination of IMAC, ion exchange and size exclusion chromatographic steps. Considering the high sequence homology and conservation of the metal ion coordinating residues of S100A8/A9 metal binding sites, the protocol is presumably applicable to S100A8/A9 of various mammalia.

Keywords: Calprotectin; IMAC (immobilized metal affinity chromatography); Nickel binding; S100A8; S100A9.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calgranulin A / genetics
  • Calgranulin A / metabolism
  • Calgranulin B* / genetics
  • Calgranulin B* / metabolism
  • Humans
  • Leukocyte L1 Antigen Complex* / metabolism
  • Sus scrofa / metabolism
  • Swine

Substances

  • Leukocyte L1 Antigen Complex
  • Calgranulin B
  • imidazoleacetic acid
  • Calgranulin A