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. © 2010 Elsevier Inc.