Standards for immunohistochemical imaging: a protein reference device for biomarker quantitation.

Academic Article

Abstract

  • We are developing a reference device to be used in the validation of immunohistochemical imaging of biomarkers by microscopy. The prototype device consists of p53 protein immobilized at various concentrations on a glass slide. The device is designed as a reference control to be used with assays that incorporate commercially available anti-p53 antibodies. p53 protein was characterized by mass spectrometry and covalently immobilized through amide linkage to the (3-aminopropyl)trietoxysilane-modified glass surface. This procedure is reproducible and provides a chemically stable product in high yield. The surface-bound protein was shown to be immunoreactive by its specific interaction with anti-p53 antibody (Ab) and detection by absorbance and fluorescence spectroscopy. Also, comparison was made with microscopic images of Ab-stained tissue samples, known to stain positive for p53. Further development will be required to establish accurate surface protein concentrations in the range required for specific clinical applications.
  • Keywords

  • Amino Acid Sequence, Biomarkers, Glass, Humans, Immobilized Proteins, Immunohistochemistry, Mass Spectrometry, Microscopy, Molecular Imaging, Quality Control, Reference Standards, Reproducibility of Results, Spectrometry, Fluorescence, Tumor Suppressor Protein p53
  • Digital Object Identifier (doi)

    Author List

  • Atha DH; Manne U; Grizzle WE; Wagner PD; Srivastava S; Reipa V
  • Start Page

  • 1005
  • End Page

  • 1014
  • Volume

  • 58
  • Issue

  • 11