Assessment of albumin removal from an immunoaffinity spin column: Critical implications for proteomic examination of the albuminome and albumin-depleted samples

Rebekah L. Gundry, Melanie Y. White, Julie Nogee, Irina Tchernyshyov, Jennifer E. Van Eyk

Research output: Contribution to journalArticle

57 Scopus citations

Abstract

High abundance proteins in serum and plasma (e.g., albumin) are routinely removed during proteomic sample processing as they can mask lower abundance proteins and peptides of biological/clinical interest. A common method of albumin depletion is based on immunoaffinity capture, and many immunoaffinity devices are designed for multiple uses. In this case, it is critical that the albumin captured on the affinity matrix is stripped from the column prior to regeneration of the matrix and processing of subsequent samples, to ensure no carryover and that maximal binding sites are available for subsequent samples. The current study examines the ability of a manufacturer's protocol to remove the proteins and peptides captured by an immunoaffinity spin column. The data presented in the current work illustrate the difficulty in completely removing albumin from the immunoaffinity device, and consequently, may explain the variability and decreased efficiency shown for this device in previous studies. In summary, the current data present important considerations for the implementation of multiple-use immunoaffinity devices for processing subsequent clinical samples in a proteomic workflow.

Original languageEnglish (US)
Pages (from-to)2021-2028
Number of pages8
JournalProteomics
Volume9
Issue number7
DOIs
StatePublished - Apr 2009

Keywords

  • Albumin
  • Antihuman serum albumin column
  • Serum

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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