Analysis of oligonucleotide photoproducts produced by UV-A light and a riboflavin photosensitizer

Stacy L. Gelhaus; William R. LaCourse

doi:10.1117/12.571348

7 December 2004 Analysis of oligonucleotide photoproducts produced by UV-A light and a riboflavin photosensitizer

Proceedings Volume 5588, Smart Medical and Biomedical Sensor Technology II; (2004) https://doi.org/10.1117/12.571348
Event: Optics East, 2004, Philadelphia, Pennsylvania, United States

Abstract

DNA damage is caused by a variety of foreign and endogenous compounds. There are endogenous photosensitizers in cells, such as porphyrins and flavins, which may create damage in the presence of UV-A light. Typically, samples are analyzed by ³²P-postlabelling and electrophoretic separation or by LC-MS separation and detection. Separation by HPLC is common; however, in all instances, the DNA sample is hydrolyzed down to nucleosides prior to analysis. It will be shown here that ion-pairing reversed phase high performance liquid chromatography (IP-RPLC) has the ability to provide biophysical information concerning the sites of UV-A induced photosensitizer damage on an intact oligonucleotide concurrent with the separation. IP-RPLC is less labor intensive and faster than electrophoretic methods and it is less costly than LC-MS. IP-RPLC can also be used to purify modified oligonucleotides for further use and analysis. This technique is sensitive to the charge, conformation, and sequence characteristics of the nucleic acid sample and may be used to determine the damage or modifications made to DNA by a variety of compounds.

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Stacy L. Gelhaus and William R. LaCourse "Analysis of oligonucleotide photoproducts produced by UV-A light and a riboflavin photosensitizer", Proc. SPIE 5588, Smart Medical and Biomedical Sensor Technology II, (7 December 2004); https://doi.org/10.1117/12.571348

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