Optimization of cardiac troponin: Pull-down by IDM affinity beads and SELDI

Diane E. Bovenkamp, Brian A. Stanley, Jennifer E. Van Eyk

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Cardiac troponin I (cTnI) is a key regulator of cardiac muscle contraction. Upon myocardial cell, injury, cTnI is lost from the cardiac myocyte and can be detected in serum, in some cases with specific disease-induced modifications, making it an important diagnostic marker for acute myocardial injury. Presently, hospital laboratories use enzyme-linked immunosorbent assays to detect cTnI, but this type of analysis lacks information about modified forms of protein (degradation or phosphorylation) that may give a more specific diagnosis from either serum or biopsies. Because cardiac and serum tissues are widely used for proteomie analysis, it is important to detect these cTnI posttranslational modifications. Therefore, we have chosen to optimize the enrichment and detection of cTnI protein by IDM Affinity Bead pull-down and surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS or SELDI) analyses. By adjusting the chemical compositions of the buffers, we have retained antibody specificity and enriched for different forms of cTnI and its associated proteins.

Original languageEnglish (US)
Title of host publicationCardiovascular Proteomics
Subtitle of host publicationMethods and Protocols
EditorsFernando Vivanco
Pages91-102
Number of pages12
DOIs
StatePublished - Feb 5 2007

Publication series

NameMethods in Molecular Biology
Volume357
ISSN (Print)1064-3745

Keywords

  • Antibody
  • Cardiac
  • IDM affinity beads
  • Mass spectrometry
  • Phosphorylation
  • Posttranslational modification
  • Purification
  • SDS-PAGE
  • SELDI
  • Serum
  • Surface-enhanced laser desorption ionization
  • Troponin
  • Western blotting
  • cTnI

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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