Hybridoma-free multiplex approach for accelerating mouse monoclonal antibody generation

DESCRIPTION provided by applicant Thanks to their exquisite specificity and sensitivity monoclonal antibodies Mab are the most important research tool for detection localization and quantification of cellular proteins and widely used for diagnosis and treatment of various human diseases Conventional hybridoma approach for generating monoclonal antibodies is time consuming labor intensive and multistep process that requires immortalization of splenocytes by fusion to a myeloma cell line partner To simplify the generation of antibodies and to increase the productivity a novel hybridoma free multiplex approach for accelerating antibody discovery is proposed The objective will be achieved by multiple antigen immunization into transgenic H Kb tsA mice harboring a mutant temperature sensitive simian virus large tumor antigen Antibody producing B cells stained with fluorescence labeled antigens will be isolated by fluorescence activation cell sorting FACS Sorted antigen specific splenocytes will be conditionally immortalized by culturing at permissive temperatures In Phase I the proposed approach will be validated by developing highly specific monoclonal antibodies to cancer related targets In Phase II the platform will be used to develop mouse monoclonal antibodies to at least protein targets that are involved in cancer Compared with traditional approach the time required for Mab generation using this approach is expected to reduce at least in half while ten fold or more increasing antibody generation productivity with significant improvement of target specificity due the selection criteria used during FACS PUBLIC HEALTH RELEVANCE As result of this project an innovative approach for generating mouse monoclonal antibodies will be developed and valuable antibody reagents for cancer research will be generated The platform will shorten time for antibody development and significantly increase productivity and antibody target specificity