Immunodepletion of albumin for two-dimensional gel detection of new mouse acute-phase protein and other plasma proteins

Xunbao Duan, David Yarmush, François Berthiaume, Arul Jayaraman, Martin L. Yarmush

Research output: Contribution to journalArticlepeer-review

37 Scopus citations

Abstract

Immunodepletion of albumin to improve the 2-D gel resolution of human plasma proteins has recently been described. With the importance of mouse models in many studies in which serum or plasma is often analyzed, we have adopted this approach to immunoprecipitate mouse albumin and evaluated its effectiveness for 2-D separation of mouse plasma proteins. Purified polyclonal antibodies against mouse albumin were effective depleting intact albumin as well as its numerous fragments from mouse plasma samples. Removal of albumin resulted in better resolution of mouse plasma proteins. Three proteins, α2- macroglobulin, coagulation factor XII, and hemopexin, that were previously either undetectable or poorly resolved, were identified from albumin-depleted 2-D gels by peptide mass fingerprinting. Albumin depletion also led to partial loss of several other proteins such as clusterin and gelsolin. This loss can be attributed to the interaction with albumin itself because the specificity of the antibody was demonstrated by Western blot. When applying this method to the 2-D separation of plasma from inflamed mouse induced by cutaneous burn injury with superimposed Pseudomonas aeruginosa infection, the upregulation of inter α-trypsin inhibitor heavy chain 4 (ITIH4) and hemopexin was unambiguously detected along with other mouse acute-phase proteins (APP), including haptoglobin and serum amyloid A. Based on the significant increase of ITIH4, we propose that this protein is a new member of mouse APP that are upregulated during the inflammatory response.

Original languageEnglish (US)
Pages (from-to)3991-4000
Number of pages10
JournalProteomics
Volume5
Issue number15
DOIs
StatePublished - Oct 2005
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Keywords

  • Acute-phase proteins
  • Albumin immunnodepletion
  • Burn and infection
  • Inter-alpha-trypsin inhibitor heavy chain 4
  • Mouse plasma 2-D gel map

Fingerprint

Dive into the research topics of 'Immunodepletion of albumin for two-dimensional gel detection of new mouse acute-phase protein and other plasma proteins'. Together they form a unique fingerprint.

Cite this