Project Number: 5030-32000-117-68-A
Project Type: Cooperative Agreement
Start Date: Dec 1, 2017
End Date: Nov 30, 2018
The objective of this research project is to develop cytokine assays for Atlantic bottlenose dolphins.
To begin with, proinflammatory molecules, as well as type II cytokine reagents will be proposed for development. These cytokines will be expressed as recombinant proteins in yeast. Expressed proteins will be purified by high performance liquid chromatography (HPLC). Bioassays will be used for testing the bioactivity of recombinant cytokines. Once bioactivity of each cytokine has been confirmed, polyclonal antibodies (pAb) to select recombinant cytokines will be produced in rabbits. Monoclonal antibodies (mAb) to specific cytokines will be produced in mice. ELISAs will be developed using the appropriate antibodies along with the appropriate recombinant cytokine. Standard curves will be used to quantify the levels of cytokines in blood or in cultures of cetacean leukocytes.