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In vivo protein biotinylation and sample preparation for the proteomic identification of organ- and disease-specific antigens accessible from the vasculature.
Journal article

In vivo protein biotinylation and sample preparation for the proteomic identification of organ- and disease-specific antigens accessible from the vasculature.

  • Roesli C Institute of Pharmaceutical Sciences, ETH Zurich, 8093 Zurich, Switzerland. christoph.roesli@pharma.ethz.ch
  • Neri D
  • Rybak JN
  • 2007-04-05
Published in:
  • Nature protocols. - 2006
Animals
Antigens
Antigens, Neoplasm
Biomarkers
Biotinylation
Blood Proteins
Mice
Organ Specificity
Perfusion
Proteomics
English Targeted delivery of bioactive molecules to diseased organs or tissues by means of binding molecules specific to markers of diseases represents a promising area of pharmaceutical intervention. The availability of markers of pathology, ideally accessible from the vasculature, is crucial for such strategies. To this aim, here we present a protocol based on terminal perfusion of mice with a reactive ester derivate of biotin that enables the covalent modification of proteins readily accessible from the bloodstream. Biotinylated proteins from total organ or tissue extracts are (i) purified on streptavidin resin in the presence of strong detergents, (ii) digested on the resin and (iii) subjected to proteomic analysis. This technology is applicable to comparative proteomic investigations of differentially expressed, accessible proteins in numerous animal models having different physiological and pathological processes.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://folia.unifr.ch/global/documents/207852
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