Development and characterization of monoclonal antibody against A disease marker protein

In this study, the production of monoclonal antibody against a disease marker selected as a targeted model, the characterization, large-scale production and purification of the produced antibodies are intended to work. For this purpose, Pancreatitis-associated protein which is a Cystic fibrosis marker used as an antigen were intraperitoneally injected to 6-8 weeks old 10 Balb/C mice at specific doses and time intervals. The antibody titers of mice were determined by indirect ELISA, and the best immune response in mice was selected for the fusion. The fusion was carried out using the mouse spleen cell and the F0 myeloma cells in the presence of polyethylene glycol (PEG). A higher immunoreactivity was detected in 3 of 10 mice used in the study. Fusion study with one of these resulted 32 hybrid clones with antibody activity. In repeated ELISA control studies, 25% of the cells with antibody activity gave a specific antibody response to the PAP antigen and 50% of them maintained this response. As a result, it has been determined that four hybridoma cells from these cells produce monoclonal antibody with high sensitivity which can be used later to develop a diagnostic kit. The obtained monoclonal antibodies were found to have a high specificity, the immunoglobulin type and the light chain type of the obtained monoclonal antibodies were determined as IgG1 and κ (kappa), respectively. Monoclonal antibodies were characterized in terms of their affinities and 2 of the fourth antibodies were found to have a high affinity. This thesis study will be an important source for future studies of the production and characterization of antibodies for the detection of animal diseases or detection of proteins for different purposes.