Student: Ali Goldensoph, Undergraduate Student in Pharmacy, Drake University
Research Mentor: Dr. Abebe Mengesha
A thin layer chromatography purification method for peptide extracts from Sample M
Radiation-induced DNA-damage represents the ultimate form of insult to the cosmonauts’ genetic material because of the chronic and heritable health effects caused by such damage. Monitoring stability of DNA during and after space travel is therefore critical. Specific proteins, including proteases are involved in DNA repair, and could serve as biomarkers of radiation-induced DNA damage. We have recently identified a novel nuclear matrix metalloprotease (nMMP) activity possibly associated with DNA repair. Over the course of the first year and 4 months of this project, our team has developed an HPLC fractionation method for analysis of bioactive peptides. Our Sample “M” was treated with dichloromethane to remove lipids and other hydrophobic molecules and extracted with 100% methanol. Sample M has been further purified using a thin layer chromatography method. The extract was then fractionated on a C-18 flash column, and the fractions were analyzed using reverse-phase HPLC. Chromatographic resolution was achieved on a C18 column using a gradient mobile phase composed of 0.1% formic acid and acetonitrile. Major component of M1 was observed at a retention time of 11.7 minutes and was quantified using VWD detector at λmax of 260 nm. In collaboration with the University of Iowa, Sample M is undergoing further molecular identification with every submission of physical Sample M from our team.