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ARS Home » Southeast Area » Auburn, Alabama » Aquatic Animal Health Research » Research » Publications at this Location » Publication #405756

Research Project: Integrated Research to Improve Aquatic Animal Health in Warmwater Aquaculture

Location: Aquatic Animal Health Research

Title: The expression of recombinant monoclonal antibodies from existing hybridoma cell lines

item CHURCHMAN, EMILY - Auburn University
item Lange, Miles
item WISE, ALLISON - Auburn University
item BRUCE, TIMOTHY - Auburn University

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 5/9/2023
Publication Date: 5/29/2023
Citation: Churchman, E.M., Lange, M.D., Wise, A.L., Bruce, T.J. 2023. The expression of recombinant monoclonal antibodies from existing hybridoma cell lines [abstract]. North American Comparative Immunology Conference Workshop, Auburn, AL. June 6-8, 2023.

Interpretive Summary:

Technical Abstract: Several monoclonal antibodies reactive to channel catfish immunoglobulins have been used for many years to aid in the study of many aspects of the functional teleost immune system. These monoclonal antibodies have traditionally originated from hybridoma technology. However, because hybridoma cell lines have the potential to lose their ability to express antibody, we have endeavored to develop these monoclonal antibodies recombinantly. We currently have a system to recombinantly express the 9E1 monoclonal antibody, which recognizes channel catfish IgM. The variable heavy and light chain domains of the 9E1 hybridoma were amplified, sequenced, and cloned into murine IgG1 and IgK expression vectors. These expression plasmids were co-transfected into 293-F cells and mature IgG was purified from culture supernatant. The recombinant 9E1 monoclonal antibody was shown to bind effectively to purified catfish IgM like the 9E1 hybridoma supernatant via dot blots. It was also demonstrated that the recombinant 9E1 monoclonal antibody binds to soluble IgM in vitro in both ELISA and ELISPOT assays. The recombinant 9E1 monoclonal antibody also has ex vivo activity to membrane-bound IgM by flow cytometry of catfish PBL and immunofluorescence with different B-cell types. Overall, the recombinant 9E1 monoclonal antibody will be a valuable tool in the examination of the channel catfish adaptive immune system. This system serves as a model for the development of additional recombinant monoclonal antibodies that will be discussed.