A recombinant 9E1 monoclonal antibody binds membrane and soluble channel catfish immunoglobulin M

Miles D Lange; Emily M Churchman; Allison L Wise; Timothy J Bruce

doi:10.1016/j.fsirep.2023.100086

A recombinant 9E1 monoclonal antibody binds membrane and soluble channel catfish immunoglobulin M

Fish Shellfish Immunol Rep. 2023 Feb 18:4:100086. doi: 10.1016/j.fsirep.2023.100086. eCollection 2023 Dec.

Authors

Miles D Lange¹, Emily M Churchman², Allison L Wise³, Timothy J Bruce³

Affiliations

¹ United States Department of Agriculture, Agricultural Research Service, Aquatic Animal Health Research Unit, Auburn, AL, USA.
² Department of Biological Sciences, Auburn University, Auburn, AL, USA.
³ School of Fisheries, Aquaculture, and Aquatic Sciences, Auburn University, Auburn, AL, USA.

Abstract

The development and validation of the recombinant 9E1 monoclonal antibody against channel catfish IgM is described. The variable heavy and light chain domains of the 9E1 hybridoma were cloned into murine IgG1 and IgK expression vectors. These expression plasmids were co-transfected into 293F cells and mature IgG was purified from culture supernatant. It is demonstrated that the recombinant 9E1 monoclonal antibody binds to soluble IgM in ELISA and ELISPOT assays and to membrane-bound IgM by immunofluorescence with different B-cell types. The recombinant 9E1 monoclonal antibody will be a valuable tool in the continued examination of the channel catfish adaptive immune system.

Keywords: Adaptive immunity; Aquaculture; Immunoassay; Reagent.