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Biochemistry
Sewell, NJ
Mentor: Lisa Ponton
Project title: Analysis of Estrogens in Wastewater Effluent with the Addition of Humic Substances
This semester was by far the most successful semester I have had with research. During the semester I finally fixed my protocols such that both the yeast reporter assay and the ELISA assay were working for my project. As for the yeast reporter assay, we finally created an appropriate estrogen solution that would dissolve in water and not crash out when added to the wells. From the yeast reporter assay we have determined that the yeast are in fact sensitive to the presence of estrogen as expected. We have also shown that a decrease in estrogen activity occurs when humic substances are present in the solution. Our hypothesis is that the humic substances bind to estrogen rendering the unable to enter into the yeast cells and cause any activity. However, we also noticed that humic substances themselves may have some estrogen activity themselves. This is an avenue that needs some more research to confirm. Overall this method was able to support our hypothesis that humic substances are in fact a positive for an environment because they decrease the activity of estrogen present in the samples.
The ELISA assay was completed to give better insight into the binding properties of estrogen and humic substances. Using an estriol ELISA assay it was noted that only a small portion of the estrogens that were added actually bound to the well walls as expected. It was also noted again that humic substances could elicit the same response as the estrogen. This may be due in part because humic substances have many polar groups that are able to interact with the same targets as estrogen. The results were even more interesting when the two compounds were incubated together. They showed increased binding to the well walls. The hypothesis for this result was that humic substances are able to orient estrogens in a favorable manner to increase their binding abilities. These results were presented at SURF this semester and my project goal was to create a lab manuscript that another student could use to continue my project with wastewater effluent rather than spiked, clean water.
Dr. Ann J. Cahill
Professor of Philosophy
Spence Pavilion 111
2340 Campus Box
Elon, NC 27244
Phone: (336) 278-5703
cahilla@elon.edu
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