Monoclonal and polyclonal antibodies against human ferritin, a nonspecific tumor marker.

Abstract

The aim of this study was to produce monoclonal and polyclonal antibodies against a nonspecific tumor marker, human ferritin. Hyperimmune ICR mice produced polyclonal antibodies after injection with 0.5 mL pristane, and were injected with NS-1 myeloma cells 2 weeks later. Hyperimmune Balb/c mice were used for the production of monoclonal antibodies (MAbs). Mice were immunized four times, given a final boost, and 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 HAT-RPMIX medium. Anti-ferritin antibody-secreting hybridoma cell lines with high titer were cloned by enzyme-linked immunoadsorbent assay (ELISA) and then subcloned by limiting dilution in 15% fetal bovine serum (FBS) HT-RPMIX medium. Five murine hybridoma-producing antiferritin MAbs were obtained and designated 1AD11F9, 1AD11E11, 2AD11D2, 2AD11A5, and 3AD11G8. Isotypes of these MAbs were identified as IgM heavy chain and kappa light chain. Hitrap Protein A and Hitrap IgM purification column were used for the purification of polyclonal and monoclonal antibodies, respectively.