High-throughput biopolymer desalting by solid-phase extraction prior to mass spectrometric analysis

被引:58
作者
Gilar, M
Belenky, A
Wang, BH
机构
[1] Waters Corp, Milford, MA 01757 USA
[2] Cetek Corp, Marlborough, MA 01752 USA
[3] Variagen Inc, Cambridge, MA 02139 USA
关键词
sample preparation; desalting; oligonucleotides; peptides;
D O I
10.1016/S0021-9673(01)00833-0
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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 (R) 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 mul) 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. (C) 2001 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:3 / 13
页数:11
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