A chip-based miniaturized format for protein-expression profiling: the exploitation of comprehensively produced antibodies

Electrophoresis. 2006 Sep;27(18):3676-83. doi: 10.1002/elps.200500821.

Abstract

Numerous antibodies have been developed and validated in recent years, and show promise for use in novel functional protein assays. Such assays would be an alternative to pre-existing comprehensive assays, such as DNA microarrays. Antibody microarrays are thought to represent those functional protein assays. While a variety of attempts have been made to apply DNA microarray technology to antibody microarrays, a fully optimized protocol has not been established. We have been conducting a project to comprehensively produce antibodies against mouse KIAA ("KI" stands for "Kazusa DNA Research Institute" and "AA" are reference characters) proteins. Using our library of antibodies, we established a novel antibody microarray format that utilizes surface plasmon resonance (SPR) technology. A label-free real-time measurement of protein expression in crude cell lysates was achieved by direct readout of the bindings using SPR. Further refinement of the antibody microarray format enabled us to detect a smaller quantity of target proteins in the lysate without the bulk effect. In this review, we first summarize available antibody array formats and then describe the above-mentioned format utilizing updated SPR technology.

Publication types

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

MeSH terms

  • Animals
  • Antibodies* / chemistry
  • Antibodies* / immunology
  • Mice
  • Miniaturization*
  • Protein Array Analysis / methods*
  • Proteins / analysis*
  • Proteins / immunology
  • Sensitivity and Specificity

Substances

  • Antibodies
  • Proteins