Aim. To generate monoclonal antibodies to Coenzyme A (CoA) with antigenic epitope specificity different to previously developed anti-CoA mAb (1F10) by introducing alternative approach for antigen preparation animal immunization. Methods. Cross-linking of CoA to Keyhole Limpet Hemocyanin (KaLH) and to Bovine serum albumin (BSA) using reversible chemical crosslinker SPDP. Hybridoma technique. Western blot analysis. Immunoprecipitation. ELISA. Results. Monoclonal antibody (A11) that specifically recognizes CoA in various immunoassays and has the same antigenic epitope specificity to existing anti-CoA mAb (1F10) has been generated. Conclusions. Application of different ways for CoA conjugation to the carrier proteins to create an antigen for immunization does not affect the antigenic epitope specificity of generated anti-CoA antibodies most probably due to the existence of only one immunogenic epitope in CoA molecule