Development and production of antibodies against gamma inactivated pathogenic bacterial spores
Ilkay Goksu Polat, Uygar Halis Tazebay, Esin Akcael- Biochemistry (medical)
- Clinical Biochemistry
- Molecular Biology
- Biochemistry
Abstract
Objectives
Gram-positive sporulated bacilli can cause many different diseases and isolation from environmental samples is difficult. Therefore, the quick detection and diagnosis of these microorganisms have critical importance because of their potentially harmful situation. However, many accepted diagnostic methods exist, and future technology points to immunoassay systems. Immunological methods to detect biological microorganisms require antigen-specific high-affinity antibodies as key materials.
Methods
In this study, Bacillus anthracis (34F2 sterne) bacterium, which causes anthrax disease, was chosen as a model organism to develop antibodies against bacterial spores. The produced spores were inactivated with gamma irradiation, and the development of monoclonal antibodies against inactivated spores was performed using hybridoma technology. Also, the polyclonal antibody was successfully obtained by immunizing the rabbit. Indirect and sandwich ELISA tests were performed to determine the antigenic properties of inactivated spores and the specific affinity of the developed antibodies.
Results
The spores, inactivated with 15 kGy, have the best-preserved surface epitopic regions and were selected as immunogen. Developed monoclonal and polyclonal antibodies were shown that there was no cross-reaction with other Bacillus species. Also, it was demonstrated that these antibodies could detect inactivated spores at a concentration of 105 spores/mL in a sandwich ELISA assay.
Conclusions
These qualified antibodies obtained will be essential in developing antibody-based diagnostic systems for spore detection from various environmental samples. This study suggests that the inactivated spores are a decent immunogen for generation antibodies and may be a candidate component for live vaccine formulation.