Different reactivities of enzyme-linked immunosorbent assays for hepatocyte growth factor

BackgroundHepatocyte growth factor (HGF) plays diverse roles in organ development, tissue regeneration, and tumor progression. Measurement of HGF concentrations in blood and tissues using enzyme-linked immunosorbent assay (ELISA) is a simple and easy way to understand the significance of HGF in tissue regeneration, pathophysiology, and diagnosis.MethodsWe evaluated 3 ELISA kits from different sources, referred to herein as kits A, B, and C for convenience.ResultsWe found that the concentrations of human HGF determined using ELISA vary significantly depending on the source of the ELISA kit. Kits A and B detected both single-chain pro-HGF and 2-chain mature HGF, but kit C detected only 2-chain HGF. A difference in reactivity was also detected during analysis of plasma samples. When rat plasma collected 4 h after subcutaneous administration of human HGF was analyzed, the HGF concentration determined using kit B was remarkably higher than those obtained using kits A and C. Results of a biological assay and Western blot analysis indicated that kit B detects even degraded HGF, by which the HGF concentrations determined using kit B were significantly overestimated.ConclusionsThis information serves as a guide for the selection of ELISA kits for human HGF.