University of Georgia

Disease: Thyroid Conditions
Sponsor: American German Shepherd Dog Charitable Foundation, the AKC Canine Health Foundation. other breed clubs
Total Grant: $97,878

Report Date: June 30, 2004

Specific Objectives 1 and 2: Amplify the canine TSHB minigene and add sequences and restriction sites to the 5’ and 3’ ends; amplify the canine a sequence and add an immunoaffinity tag on the 3’ end. These steps have been completed.

Specific Objectives 3 and 5: Produce a yoke canine TSH for insertion into the PEAK expression system and express the protein in human embryonic kidney (HEK) cells. These steps have been postponed with efforts directed into cloning sequences into the pBud expression system.

Specific Objective 4: Co-express cTSHB and the a subunit in HEK cells. This step is being developed using pBud vectors. Specific Objective 6: Purify recombinant cTSH using immunoaffinity columns. This step has begun with efforts currently to increase the expression level by order of magnitude. Specific Objectives 7 through 11: Assess cross-reactivity with various monoclonal antibodies, assess glycosylation pattern, prepare poyclonal antibody serum, and purify and label the IgG for use in detection systems (tests). Some initial work has begun, but primary efforts await expression and purification of rcTSH 9 steps 4 and 6).

Grant Results for Thyroid Conditions - Complete Report - 06/30/2004

Grant Results for Thyroid Conditions - Complete Report - 12/09/2004