The aim of this study was to produce monoclonal and polyclonal antibodies against heart-type fatty acid-binding protein (HFABP). Hyperimmune ICR mice produced polyclonal antibodies after injection with 0.5 mL pristane, and were injected with NS-1 myeloma cells two weeks later. Hyperimmune Balb/c mice were used for the production of monoclonal antibodies (MAbs). After these mice were immunized four times and given a final boost, their spleen cells were collected and fused with NS-1 myeloma cells under the presence of PEG 1500. The fused cells were then selected in the hypoxathine, aminopterine, and thymidine (HAT)-RPMIX medium. Anti- HFABP antibody-secreting hybridoma cell lines with high titer were cloned by enzyme-linked immunosorbent assay (ELISA) and then subcloned by limiting dilution in 15% fetal bovine serum (FBS) HT-RPMIX medium. Nine murine hybridoma producing anti- HFABP MAbs were obtained and designated HB1, HC2, HE4, HB9, HA2, HC7, HD6, HD11, and HE5. Isotypes of these MAbs were identified as IgG2a heavy chain and κ light chain. Hitrap Protein A column was used for the purification of polyclonal and monoclonal antibodies.
