Characterization of Carp (Cyprinus carpio L.) Immunoglobulin Structure

Choi, Sang-Hoon; Park, Kwan-Ha; Yoon, Jong-Man

doi:2002.15.2.290

Article

Animal Breeding and Genetics

Asian-Australasian Journal of Animal Sciences 2002;15(2): 290-296.
https://doi.org/10.5713/ajas.2002.290 Published online January 1, 2002.

Characterization of Carp (Cyprinus carpio L.) Immunoglobulin Structure

Sang-Hoon Choi, Kwan-Ha Park, Jong-Man Yoon

Abstract

Serum immunoglobulins (Igs) from Israeli carp were purified using affinity chromatography. Fish were immunized with purified mouse IgG, and the specific fish antibodies were purified from the immune serum on a mouse IgG-immobilized agarose gel. Rabbit anti-Israeli carp Igs (R I. carp Igs) antibodies were produced following hyperimmunization with mouse IgG specific carp antibodies. SDS-PAGE analysis under reducing condition showed that Israeli carp Igs were composed of two -like heavy chains with about 82 and 50 kd, respectively, and one light chain with about 25 kd. On immunoblotting analysis, however, R I. carp Igs failed to react with the light chain. When both protein A and protein G-purified normal carp Ig were compared with mouse IgG-specific Israeli carp Ig, no significant structural differences among them were observed. To investigate if there is any homology between other fish Ig molecules, cross-reactivity of R I. carp Igs against Ig molecules from 6 different fish sera and mouse control serum was checked on immunoblotting analysis. As a result, R I. carp Igs responded to Israeli carp, common carp, and tilapia Ig molecules. In flow cytometry study, however, R I. carp Igs appeared to recognize 42.0%, 35.8% and <5% of Israeli carp, common carp and tilapia Ig+ head kidney cells, respectively. The result suggests the heterogeneity between receptor Igs on B-like lymphocytes and soluble Igs in serum. It is crucial to obtain pure fish Igs to produce reagent antibodies as tools for the study on their specific immune responses.

Keywords: Affinity Chromatography; Flow Cytometry; Carp Ig; Tilapia