High-throughput biopolymer desalting by solid-phase extraction prior to mass spectrometric analysis.
In the last 10 years mass spectrometry (MS) has become an important method for analysis of peptides, proteins and DNA. It was recently utilized for accurate high-throughput protein identification, sequencing and DNA genotyping. The presence of non-volatile buffers compromises sensitivity and accuracy of MS biopolymer analysis; it is essential to remove sample contaminants prior to analysis. We have developed a fast and efficient method for desalting of DNA oligonucleotides and peptides using 96-well solid-phase extraction plates packed with 5 mg of Waters Oasis HLB sorbent (Waters, Milford, MA, USA). This reversed-phase sorbent retains the biopolymer analytes, while non-retained inorganic ions are washed out with pure deionized water. DNA oligonucleotides or peptides are eluted using a small amount (20-100 microl) of acetonitrile-water (70:30, v/v) solution. The SPE desalting performance meets the requirements for MS applications such as protein digest analysis and DNA genotyping.
Gilar
M
M
Waters Corporation, Milford, MA 01757-3696, USA. martin_gilar@waters.com
Belenky
A
A
Wang
B H
BH
D001704:Biopolymers / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D003574:Cytochrome c Group / Q000737:chemistry; D004247:DNA / Q000302:isolation & purification*; D010449:Peptide Mapping; D012492:Salts; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D001704:Biopolymers; D003574:Cytochrome c Group; D012492:Salts; D004247:DNA