Burbot is the only freshwater member of the cod-like fish (Gadiformes) and are an ideal candidate for cool/cold-water aquaculture. Burbot are susceptible to disease outbreaks, and may become asymptomatic carriers, however they are refractory or even show limited mortality to most salmonid pathogens. However, larval and juvenile burbot have been shown to be susceptible to Aeromonas species, with mortality from ranging from 10 to 20% following laboratory challenges. Outbreaks of a species of Aeromonas most closely related to A. veronii have been observed to cause mortality up to 92% in juveniles. Development and optimization of a serological method using monoclonal antibodies to survey juvenile burbot for Aeromonas species, and aid in the generation of a vaccine, is paramount to the success of burbot aquaculture.
IgM proteins were isolated from burbot serum via an agarose affinity column containing immobilized mannan binding protein. The purified burbot IgM was injected into three mice for the generation of antibodies demonstrating specific affinity to burbot IgM, determined via ELISA (enzyme linked immunosorbent assay). Mice possessing antibodies for burbot IgM were culled and the spleens harvested to generate hybridoma cell lines. Using an ELISA the hybridoma cell lines were screened against the heavy and light chains of burbot IgM to identify a candidate for an anti-burbot IgM monoclonal antibody. Four hybridoma cell lines yielded specificity to burbot IgM heavy and or light chain. Three cell lines had affinity for the heavy chain, and two showed affinity to the light chain. The cell line showing the highest absorbance for burbot heavy chain is currently being used to develop and optimize an ELISA to measure immune response of burbot to Aeromonas infection. This optimized ELISA will serve as a primary tool to aid in the development of a vaccine to prevent Aeromonas species infections in burbot aquaculture.