Submitted to: Veterinary Immunology and Immunopathology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: December 20, 2002
Publication Date: February 10, 2003
Citation: Shelby, R.A., Shoemaker, C.A., Klesius, P.H. 2003. Development of an Immunoassay to measure the humoral immune response of hybrid striped bass Morone chrysops X M. saxatilis. Veterinary Immunology and Immunopathology. 91(3-4):217-225. Interpretive Summary: This paper describes the chemical properties of immunoglobulin (Ig) protein found in the blood of hybrid striped bass. one polyclonal and two monoclonal antibodies were developed which are specific for the Ig. They were used in the enzyme-linked immunosorbent assay (ELISA) test to quantitatively determine the amount of circulating antibody in the fish. This is important because levels of Ig reflect the immune status of the fish with respect to specific disease organisms, and also the general health of the animal. Using the ELISA test, the kinetics of the immune response of these fish was followed for a period of 98 days following administration of an antigen. The ELISA will be used in future experiments to monitor the immune response of hybrid striped bass to vaccination and disease.
Technical Abstract: Hybrid striped bass (HSB) were immunized with bovine serum albumin (BSA) and the specific anti-BSA immunoglobulin (Ig) was affinity purified from the resulting serum by means of an agarose gel-BSA column. The native Ig had an apparent molecular size of 893 kDa, by size exclusion chromatography, and when examined by polyacrylamide gel electrophoresis under denaturing conditions, resolved to heavy (H) and light (L) chains of 76 and 27 kDa. Affinity purified native HSB Ig was used to immunize a goat which produced specific anti-HSB Ig antibody (Ab). Purified native HSB Ig was also used to produce two murine monoclonal antibodies (mAbs) with specific affinities for H and L chain moieties of the HSB Ig molecule. Both polyclonal and monoclonal antibodies could be used individually in an indirect enzyme- linked immunosorbent assay (ELISA) to measure specific anti-BSA Ig in HSB serum. These antibodies could also be used in combination to measure total Ig in a capture ELISA format. Using both assays, the kinetics of the humoral immune response of HSB was measured for 98 days following two injections of BSA.