MALDI-target integrated platform for affinity-captured protein digestion

Anal Chim Acta. 2014 Jan 7:807:1-8. doi: 10.1016/j.aca.2013.08.051. Epub 2013 Sep 11.

Abstract

To address immunocapture of proteins in large cohorts of clinical samples high throughput sample processing is required. Here a method using the proteomic sample platform, ISET (integrated selective enrichment target) that integrates highly specific immunoaffinity capture of protein biomarker, digestion and sample cleanup with a direct interface to mass spectrometry is presented. The robustness of the on-ISET protein digestion protocol was validated by MALDI MS analysis of model proteins, ranging from 40 fmol to 1 pmol per nanovial. On-ISET digestion and MALDI MS/MS analysis of immunoaffinity captured disease-associated biomarker PSA (prostate specific antigen) from human seminal plasma are presented.

Keywords: Digestion; ISET; Immunoaffinity; MALDI; MRM; MS; PSA; Proteomics; RP; SPE; SPR; integrated selective enrichment target; mass spectrometry; matrix-assisted laser desorption/ionization; multiple reaction monitoring; prostate specific antigen; reverse phase; solid-phase extraction; surface plasmon resonance.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkylation
  • Antibodies, Monoclonal / immunology
  • Chromatography, Affinity
  • Humans
  • Male
  • Oxidation-Reduction
  • Prostate-Specific Antigen / blood*
  • Prostate-Specific Antigen / immunology
  • Prostate-Specific Antigen / isolation & purification
  • Proteomics
  • Solid Phase Extraction
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization*

Substances

  • Antibodies, Monoclonal
  • Prostate-Specific Antigen