A general method for producing bioaffinity MALDI probes.
A bioaffinity probe based on the idea of immobilizing avidin on the probe surface to extract biotinylated oligosaccharide is described. The probe is produced by taking advantage of the natural affinity of proteins for hydrophobic polymer films. The avidin is immobilized by simply drying the solution on a polymer film surface. This produces a bioaffinity probe that shows enhanced activity for biotin-labeled oligosaccharides. The probe is produced in a matter of minutes but is highly effective for concentrating biotinylated oligosaccharide on the surface. The best matrix for the analysis is DHB, and the best film for the probe is a polyester material commonly used for transparency film. The efficacy of the probe is illustrated with neutral and anionic oligosaccharides. Oligosaccharides derivatized with biotin are retained while those that are unlabeled are washed away. No trace of the unlabeled oligosaccharide is observed in the mass spectrum.
Wang
H
H
Department of Chemistry, University of California, Davis 95616, USA.
Tseng
K
K
Lebrilla
C B
CB
D000345:Affinity Labels / Q000138:chemical synthesis*; D001710:Biotin / Q000138:chemical synthesis; D002240:Carbohydrate Sequence; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000138:chemical synthesis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D000345:Affinity Labels; D009844:Oligosaccharides; D001710:Biotin