Analysis of Protein Expression in Cell Microarrays: A Tool for Antibody-Based Proteomics

J Histochem Cytochem. 2006 Dec;54(12):1413-23. doi: 10.1369/jhc.6A7001.2006. Epub 2006 Sep 6.

Abstract

Tissue microarray (TMA) technology provides a possibility to explore protein expression patterns in a multitude of normal and disease tissues in a high-throughput setting. Although TMAs have been used for analysis of tissue samples, robust methods for studying in vitro cultured cell lines and cell aspirates in a TMA format have been lacking. We have adopted a technique to homogeneously distribute cells in an agarose gel matrix, creating an artificial tissue. This enables simultaneous profiling of protein expression in suspension- and adherent-grown cell samples assembled in a microarray. In addition, the present study provides an optimized strategy for the basic laboratory steps to efficiently produce TMAs. Presented modifications resulted in an improved quality of specimens and a higher section yield compared with standard TMA production protocols. Sections from the generated cell TMAs were tested for immunohistochemical staining properties using 20 well-characterized antibodies. Comparison of immunoreactivity in cultured dispersed cells and corresponding cells in tissue samples showed congruent results for all tested antibodies. We conclude that a modified TMA technique, including cell samples, provides a valuable tool for high-throughput analysis of protein expression, and that this technique can be used for global approaches to explore the human proteome.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies / immunology*
  • Antigen-Antibody Reactions
  • Cell Line
  • Cells, Cultured
  • Electrophoresis, Agar Gel
  • Female
  • Humans
  • Immunohistochemistry
  • Leukocytes, Mononuclear / immunology
  • Male
  • Proteins / analysis*
  • Proteins / genetics*
  • Proteins / immunology
  • Proteomics*
  • Sensitivity and Specificity
  • Tissue Array Analysis / methods*

Substances

  • Antibodies
  • Proteins