The aim of this study was to produce monoclonal and polyclonal antibodies against C-reactive protein (CRP). 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-CRP 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. Ten murine hybridoma producing anti- CRP MAbs were obtained and designated CRP-2E, CRP-2B, CRP-5E, CRP-11H, CRP-11F, CRP-12C, CRP-3C, CRP-12F, CRP-11G, CRP-9G. Aside from the isotypes of CRP-3C, CRP-12C and CRP-5E were identified as IgA and κ light chain, those of the others were identified as IgG2b heavy chain and κ light chain. Hitrap rProtein A column was used for the purification of polyclonal and monoclonal antibodies.
