Exploring salivary glucosyltransferase B as a possible marker for caries activity
Stockton, Jonathan B.
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Objectives . The ultimate goal of this project is to develop a chairside dental caries diagnostic test that will quantitate Streptococcus mutans glucosyltransferase B in saliva to identify patients with high caries risk. Full length GtfB and three fragments (N-terminal 1/3, N-terminal 2/3 and C-terminal 1/3) were expressed in Escherichia coli , purified and used to immunize mice, from which 100 IgG monoclonal antibodies were screened for reactivity to Gtf-B. Results . Characterization and selection of these antibodies included identifying the following properties; (1) reactivity with native GtfB from S. mutans culture supernatants (immunoblotting); (2) and ability to bind to native GtfB with high affinity (as measured by immunoprecipitation). Initial screening revealed 12 antibodies with strong reactivity. Saliva samples were collected and the Gtf-B was measured in each sample with a combination of 4 antibodies in an ELISA format. Finally, each antibody was screened for inhibitory effect on Glucosyltransferase activity. Conclusion . Our data showed no relationship between S. mutans counts and DMFT/DMFS scores. Furthermore, our Gtf-B scores showed an inverse relationship when compared to active decay lesions. When the antibodies were screened for inhibitory effect one, 11E10, showed strong inhibitory activity (no statistical analysis).