| Abstract [eng] |
Antibiotic resistance is one of the biggest problems healthcare systems face in the 21st century. Although the development of antibiotics during the last 100 years has significantly decreased the load of infectious diseases, they are on the rise again as poor regulation of antibiotic use is causing the development of antimicrobial resistance. The lack of affordable and rapid diagnostics challenges the correct prescription of antibiotics. Immunodetection is a specific, sensitive, and cost-effective approach to detect antibiotic resistance proteins. However, the development of such diagnostic tests is limited by the lack of monoclonal antibodies against antibiotic resistance proteins. The aim of this work was to generate mouse monoclonal antibodies against OXA-48, OXA-134, SME-3, SHV-42, and ADC-144 proteins which confer resistance to various types of β-lactam antibiotics. Monoclonal antibodies (MAbs) were produced by hybridoma technology. Initially, OXA-48 and OXA-134 β-lactamases were purified, all β-lactamases were used as immunogens and thirteen MAb secreting cell lines were generated. MAbs were then characterized by immunochemical methods. It was shown that eleven out of thirteen MAbs exhibit high affinity towards their respective antigens, and twelve of all the antibodies recognize linear epitopes. Furthermore, sandwich ELISA assays were developed detecting three different recombinant β-lactamases. Additional testing is needed to assess the MAbs true diagnostic potential. |
