Preparation and Characterization of Monoclonal Antibody Against Truncated Recombinant Nuclear Matrix Protein (NMP22)

Background: Bladder cancer is a major worldwide health problem. Diagnosis of acute and chronic bladder carcinoma is based on the detection of a number of tumor markers such as nmp22 (nuclear matrix protein 22). Nmp22 is one of the tumor markers used for detecting the recurrence of bladder cancer. Objectives: The aim of this study was to develop hybrid cell producing monoclonal antibodies (MAbs), specifically for nmp22 using hybridoma technology. Materials and Methods: Complete and incomplete adjuvant with recombinant truncated nmp22 antigens were emulsified and injected to BALB/c mice. The spleen was removed and the splenocytes were fused with sp2/0 myeloma cells. Characterization of the produced mAb was carried out. Results: As a result of the fusion, hybridoma cells were produced and exhibited high- titer antibodies. By testing of colons based on the EnzymeLinkedImmunosorbent Assay (ELISA), 135hybridomacolonswere selected, out of whicheight high titer clones including 54D-6F-2E-36-7H-2A-8E-1D-6D were detected and one of the clones named FPR92 was selected, and the produced mAb was further characterized. The produced antibody belongs to the IgG class and its light chain was kappa. With respect to affinity, the mAb was included as high affinity 3�10-7 reacting with NMP22 recombinant protein .The western blot of cancerous bladder tissue showed the presence of 40 and 55 kD proteins as major bands that reacted with this mAb. Conclusions: Based on a double limiting dilution protocol, a type of monoclonal antibody, named FPR92 was produced and characterized. Keywords: Nmp22, Hybridoma, mAb, FPR92, Characterization