Monoclonal and polyclonal antibodies against human ferritin, a nonspecific tumor marker.
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.
Chou
S F
SF
Department of Agricultural Chemistry, National Taiwan University, Taipei, ROC.
Chen
C Y
CY
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D014408:Biomarkers, Tumor / Q000276:immunology*; D015374:Biosensing Techniques; D002846:Chromatography, Affinity; D005293:Ferritins / Q000276:immunology*; D006801:Humans; D006825:Hybridomas; D007143:Immunoglobulin Heavy Chains / Q000276:immunology; D007132:Immunoglobulin Isotypes / Q000276:immunology; D007075:Immunoglobulin M / Q000276:immunology; D007145:Immunoglobulin kappa-Chains / Q000276:immunology; D051379:Mice; D008813:Mice, Inbred ICR
D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor; D007143:Immunoglobulin Heavy Chains; D007132:Immunoglobulin Isotypes; D007075:Immunoglobulin M; D007145:Immunoglobulin kappa-Chains; D005293:Ferritins