Quantitative Analysis of Oligonucleotides by Matrix-assisted Laser Desorption/Ionization Mass Spectrometry

✍ Scribed by Bernd A. Bruenner; Tai-Tung Yip; T. William Hutchens

• Publisher: John Wiley and Sons
• Year: 1996
• Tongue: English
• Weight: 434 KB
• Volume: 10
• Category: Article
• ISSN: 0951-4198
• DOI: 10.1002/(sici)1097-0231(199611)10:14<1797::aid-rcm754>3.0.co;2-5

✦ Synopsis

Quantitative aspects of oligonucleotide analysis by matrix-assisted laser desorption/iotion (MALDI) mass spectrometry remain largely unexplored relative to the efforts that have been devoted to quantitative peptide and protein analysis. The successful quantitation of these other biopolymers coupled with the potential of rapid nucleic acid analysis by desorptionhonization techniques prompted the present investigation into quantifying mixed base oligonucleotides of intermediate molecular weights. This report describes the concentrationdependent desorptionlionization of a 21-base oligonucleotide (MW 6361) using a %-base oligonucleotide (MW 11 131) as an internal standard. Peak height and peak area ratios ( d y t e to internal standard) varied linearly as a function of oligonucleotide concentration (R2=0.966 and 0.991, respectively). The linearity of response extended over nearly three orders of magnitude, from 0.125 to 100 pmol of analyte applied. The use of an internal standard improved the linearity of the calibration curve and reduced relative standard deviations. These results demonstrate for the first time the quantitation of medium size oligonucleotides using MALDI.