Investigation of an albumin-enriched fraction of human serum and its albuminome

Rebekah L. Gundry, Qin Fu, Christine A. Jelinek, Jennifer E. Van Eyk, Robert J. Cotter

Research output: Contribution to journalArticle

Abstract

The removal of albumin and other high abundance proteins is a routine first step in the analysis of serum and plasma proteomes. However, as albumin can bind proteins and peptides, there is a universal concern as to how the serum proteome is changed by the removal of albumin. To address this concern, the current study was designed to identify proteins and peptides removed from the serum during albumin depletion; to determine which of these are bound to albumin (rather than copurified) and whether the bound proteins are intact proteins or peptide fragments. Sequential, independent analyses including both anti-albumin antibody (anti-HSA) affinity chromatography and SEC were used to isolate albumin-bound proteins. RP-HPLC and 1-D SDS-PAGE were then used to further separate the proteins prior to identification by MS/MS. Finally, whole protein molecular weight (MW) MS measurements coupled with protein coverage obtained by MS were combined to assess whether the bound proteins were intact or peptide fragments. Combining the results from multiple approaches, 35 proteins, of which 24 are intact, were found to be associated with albumin, and they include both known high and low abundance proteins.

Original languageEnglish (US)
Pages (from-to)73-88
Number of pages16
JournalProteomics - Clinical Applications
Volume1
Issue number1
DOIs
StatePublished - Jan 2007

Fingerprint

Albumins
Serum
Proteins
Peptide Fragments
Proteome
Affinity chromatography
Peptides
Weighing
Affinity Chromatography
Serum Albumin
Polyacrylamide Gel Electrophoresis
Anti-Idiotypic Antibodies
Molecular Weight
Molecular weight
High Pressure Liquid Chromatography
Plasmas

Keywords

  • Albumin depletion
  • Albumin-binding proteins
  • Albuminome
  • Anti-HSA
  • SEC

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Investigation of an albumin-enriched fraction of human serum and its albuminome. / Gundry, Rebekah L.; Fu, Qin; Jelinek, Christine A.; Van Eyk, Jennifer E.; Cotter, Robert J.

In: Proteomics - Clinical Applications, Vol. 1, No. 1, 01.2007, p. 73-88.

Research output: Contribution to journalArticle

Gundry, Rebekah L. ; Fu, Qin ; Jelinek, Christine A. ; Van Eyk, Jennifer E. ; Cotter, Robert J. / Investigation of an albumin-enriched fraction of human serum and its albuminome. In: Proteomics - Clinical Applications. 2007 ; Vol. 1, No. 1. pp. 73-88.
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