Highly sensitive recombinant antibodies capable of reliably differentiating heart-type fatty acid binding protein from noncardiac isoforms

Anal Biochem. 2010 Dec 15;407(2):165-71. doi: 10.1016/j.ab.2010.07.033. Epub 2010 Aug 7.

Abstract

During recent times, heart-type fatty acid binding protein (hFABP) has gained increasing credence as a promising cardiac biomarker. This is largely due to its rapid myocardial release and subsequent clearance kinetics, which are superior to those of myoglobin and offer an earlier diagnostic window than the troponins. Realization of its full diagnostic and prognostic potential is dependent on accessibility to robust hFABP-specific assays. Here we describe a rational strategy for generation and screening of hFABP-specific avian-derived recombinant antibodies. These antibodies were confirmed to be exquisitely specific for hFABP, with no cross-reactivity observed in a representative panel of the most homologous non-heart-type FABP isoforms. All of the antibodies tested exhibited single-figure nanomolar affinities, and their analytical potential was demonstrated in a simple inhibition enzyme-linked immunosorbent assay (ELISA) format that returned an impressive limit of quantitation (LOQ) value of 1.9 ng/ml. The cumulative results underline the potential value of these antibodies as enabling reagents for use in a variety of immunodiagnostic configurations.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cross Reactions
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Fatty Acid Binding Protein 3
  • Fatty Acid-Binding Proteins / analysis*
  • Fatty Acid-Binding Proteins / chemistry
  • Fatty Acid-Binding Proteins / immunology
  • Humans
  • Kinetics
  • Myocardium / metabolism
  • Myoglobin / chemistry
  • Peptide Library
  • Protein Isoforms / analysis
  • Protein Isoforms / chemistry
  • Protein Isoforms / immunology
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / immunology
  • Recombinant Proteins / metabolism
  • Single-Chain Antibodies / chemistry
  • Single-Chain Antibodies / genetics
  • Single-Chain Antibodies / immunology*
  • Surface Plasmon Resonance

Substances

  • FABP3 protein, human
  • Fatty Acid Binding Protein 3
  • Fatty Acid-Binding Proteins
  • Myoglobin
  • Peptide Library
  • Protein Isoforms
  • Recombinant Proteins
  • Single-Chain Antibodies