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On-plate selective enrichment and self-desalting of peptides/proteins for direct MALDI MS analysis.

Analytical chemistry (2012-02-14)
Zhoufang Zeng, Yandong Wang, Shoulei Shi, Lifeng Wang, Xinhua Guo, Nan Lu
ABSTRACT

In this paper, a new technique has been proposed to achieve simultaneous peptides/proteins enrichment and wash-free self-desalting on a novel sample support with a circle hydrophobic-hydrophilic-hydrophobic pattern. Upon deposition, the sample solution is first concentrated in a small area by repulsion of the hydrophobic outer layer, and then, the peptides/proteins and coexisting salt contaminants are selectively captured in different regions of the pattern through strong hydrophobic and hydrophilic attractions, respectively. As a result, the detection sensitivity is improved by 2 orders of magnitude better than the use of the traditional MALDI plate, and high-quality mass spectra are obtained even in the presence of NaCl (1 M), NH(4)HCO(3) (100 mM), or urea (1 M). The practical application of this method is further demonstrated by the successful analysis of myoglobin digests with high sequence coverage, demonstrating the great potential in proteomic research.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ammonium bicarbonate, BioUltra, ≥99.5% (T)
Sigma-Aldrich
Ammonium bicarbonate, ReagentPlus®, ≥99.0%