Integrated Approach to Desalting and Concetrating Protein/Peptide Samples for MALDI MS Analysis.

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Weibin Chen;Peter J. Lee;Edouard S. P. Bouvier;Jeffrey W. Finch, John C. Gebler;Jeff Brown;Emmanuelle Claude;Dominic Gostick;James Langridge
Fall ACS 2002; 224th ACS National Meeting; Boston August 18-22
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Content Subtype:
Protein; Peptide; Ovalbumin
Methods for preparation/purification of peptide/protein samples often result in an overwhelming stoichiometric excess of inorganic salts and detergents that interfere with MALDI analysis. A clean-up step following sample preparation is frequently necessary for a successful MADLI analysis. Desalting of the samples is often time consuming and leads to sample loss. An innovative sample preparation technology for peptide and protein analysis via MALDI mass spectrometry is described in this presentation. The technology utilizes a MALDI plate as a platform, enabling on-target cleanup and high-volume sample loading, considerably simplifying sample preparation process. The presentation will demonstrate that contaminated peptide/protein samples can be directly analyzed by a MALDI mass spectrometer after on-target cleanup. Examples with commonly used salts or detergents will show that the cleanup does not affect the overall sensitivity and is as least as efficient as other common purification methods. In addition, the design allows the use of an increased sample volume so samples with low analyte concentrations can be focused to a confined region. Analysis of peptide mixtures (artificial mixture and protein digests) performed bythe plate shows excellent sensitivity typically one order magnitude improvement over standard stainless steel plates).

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