| Abstract [eng] |
Monoclonal and recombinant antibodies (MAbs and RAbs) specifically recognize antigens and are widely used in research, diagnostics and therapy. They can target carbonic anhydrase IX and XII (CA IX and CA XII), which are considered as promising cancer biomarkers. These isozymes are involved in carcinogenesis by contributing to extracellular pH regulation under hypoxia conditions, thus promoting tumor cell growth and survival. The newly developed MAbs can be exploited for antibody-based CA IX and CA XII expression studies and cancer diagnostics. Antigen-specific RAbs are less immunogenic, thus can be used as immunotherapeutic agents. In the current study a novel collection of MAbs against recombinant CA IX and CA XII was developed for studying expression of CA IX and CA XII in biological specimens. The MAbs were evaluated in different immunoassays – their cross-reactivity, affinity, neutralizing activity was investigated. The CA XII-specific MAb 14D6 with a great potential for diagnostics and therapy was selected, which recognized cellular CA XII on the surface of live cancer cells and blocked the catalytic activity of recombinant CA XII. We have selected this MAb for the generation of two RAb variants: single chain fragment variable (scFv) and scFv fused with human immunoglobulin Fc fragment (scFv-Fc). Both RAbs specifically recognized recombinant CA XII thus demonstrating their potential for developing therapeutic agents. |
