A protein microarray ELISA for screening biological fluids

Methods Mol Biol. 2004:264:161-72. doi: 10.1385/1-59259-759-9:161.

Abstract

Protein microarrays permit the simultaneous measurement of many proteins in a small sample volume and therefore provide an attractive approach for the quantitative measurement of proteins in biological fluids, including serum. This chapter describes a microarray enzyme-linked immunosorbent assay (ELISA). Capture antibodies are immobilized onto a glass surface; the covalently attached antibodies bind a specific antigen from a sample overlaying the array. A second, biotinylated antibody that recognizes the same antigen as the first antibody, but at a different epitope, is then used for detection. Detection is based on an enzymatic signal-enhancement method known as tyramide signal amplification (TSA). By coupling a microarray-ELISA format with the signal amplification of tyramide deposition, the assay sensitivity is as low as sub-pg/mL.

Publication types

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

MeSH terms

  • Amides / chemistry
  • Amides / metabolism
  • Antibodies / metabolism
  • Antibody Affinity
  • Biotin / metabolism
  • Body Fluids / chemistry*
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Epitopes
  • Protein Array Analysis / methods*
  • Proteins / analysis*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Streptavidin / metabolism

Substances

  • Amides
  • Antibodies
  • Epitopes
  • Proteins
  • Biotin
  • Streptavidin