Isolation of Immunoglobulin G from the Atlantic Bottlenose Dolphin, (Tursiops truncatus)
IAAAM 1999
Kathleen E. Kohlberg1, BS; Karen G. Burnett2, PhD
1Grice Marine Lab, University of Charleston SC, Charleston, SC, USA; 2Department of Microbiology and Immunology, Medical University of South Carolina, Charleston, SC, USA


Immunoglobulin G (IgG) is the dominant antibody in the vertebrate peripheral immune system. Although well studied in land mammals, its characteristics are not well understood in marine mammals such as the Atlantic bottlenose dolphin, Tursiops truncatus. The present research has focused on the evaluation of several methods for the isolation of dolphin IgG, with the goal of facilitating studies of the immunocompetence of T. truncatus. The most successful method to date has been FPLC (Fast Protein Liquid Chromatography) using a Protein G Sepharose 4 Fast Flow column. Serum obtained from wild animals off the coast of Sarasota, FL was centrifuged, filtered, then run over the column. Proteins were eluted off the column by switching to a glycine buffer. One large initial peak was observed, representing all of the serum proteins and other components that did not bind Protein G. After the addition of glycine, a second, smaller peak was observable, presumably IgG. Further experiments, including SDS-PAGE and Western blot analysis, need to be conducted on these samples to confirm that it is dolphin IgG. Once IgG has been purified, it will be used to create a monoclonal antibody that will be a tool for health assessment in these animals, by measuring amounts of dolphin IgG in serum and enumerating B cells.


This work was supported by a Slocum-Lunz supply grant. The authors wish to thank Rhonda Patterson, Ph.D., Gus DiNovo, M.S., Philippe Arnaud, Ph.D., and Kent MacDougal for technical assistance.

Speaker Information
(click the speaker's name to view other papers and abstracts submitted by this speaker)

Kathleen E. Kohlberg
Grice Marine Lab, University of Charleston SC
Charleston, SC, USA

MAIN : Session II : Isolation of IgG
Powered By VIN