Determination of a vascular endothelial growth factor - aptamer photo-crosslink

ANYL 94

Irena Nikcevic, nikcevi@email.uc.edu1, Patrick A Limbach, Pat.Limbach@uc.edu1, Kenneth R. Wehmeyer, wehmeyer.kr@pg.com2, William R. Heineman, heinemwr@ucmail.uc.edu1, and Carl J. Seliskar, carl.seliskar@uc.edu1. (1) Department of Chemistry, University of Cincinnati, P.O.Box 210172, Cincinnati, OH 45221, (2) Analytical Sciences Department, Procter & Gamble Pharmaceuticals, 8700 Mason-Montgomery Rd, Mason, OH 45040
The objective of this work was to explore the potential of photoSELEX to identify high-affinity binding, specifically the photo-crosslinking of a fluorescently labeled photoaptamer to the vascular endothelial growth factor (VEGF) target. Photoaptamers can provide an increased level of selectivity due to the spatial requirements for crosslinking. Brief exposure to a UV light creates covalent crosslinks between the modified nucleotide and amino acids of a protein or polypeptide target. The aptamer/protein ratio was optimized in the photo-crosslinking reactions for maximum crosslinking. After incubation at room temperature for 15 min, each reaction mixture was exposed to UV light at 313 nm and 3mJ/cm2. The result obtained was identified using 3% agarose and a 12% PAGE gel mobility shift assay, by positive ion electrospray ionization mass spectrometry and microchip capillary electrophoresis.
 

General Posters
7:00 PM-9:00 PM, Sunday, August 19, 2007 BCEC -- Exhibit Hall - B2, Poster

Sci-Mix
8:00 PM-10:00 PM, Monday, August 20, 2007 BCEC -- Exhibit Hall - B2, Sci-Mix

Division of Analytical Chemistry

The 234th ACS National Meeting, Boston, MA, August 19-23, 2007