Salivary protein/peptide profiling with SELDI-TOF-MS.
Raymond Schipper1, Arnoud Loof, Jolan de Groot
1Wageningen Centre for Food Sciences, TI Food and Nutrition, Laboratory of Food Chemistry, Wageningen University, the Netherlands. raymond.schipper@wur.nl
Annals of the New York Academy of Sciences
|April 17, 2007
Summary
Surface-enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS) effectively profiles low-mass saliva proteins. Factors like chip surface, sample type, and storage temperature significantly impact salivary proteome analysis.
Area of Science:
- Proteomics
- Biochemistry
- Analytical Chemistry
Background:
- The human saliva proteome is complex and dynamic.
- Understanding salivary protein composition is crucial for diagnostics and understanding physiological processes.
- High-throughput methods are needed for comprehensive salivary proteome analysis.
Purpose of the Study:
- To demonstrate large-scale profiling of salivary proteins and peptides using SELDI-TOF-MS.
- To investigate the impact of pre-analytical variables on saliva proteome analysis.
- To establish SELDI-TOF-MS as a tool for studying saliva proteome dynamics.
Main Methods:
- Surface-enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS) was employed for protein profiling.
- Saliva samples were analyzed to detect proteins and peptides in the 2 to 100 kDa range.
- The influence of chip surface type, sample type, processing time, storage temperature, centrifugation, and freeze-thaw cycles was evaluated.
Main Results:
- Chip surface and sample type significantly influenced the detected salivary protein amount and composition.
- Delayed sample processing led to proteolysis, altering peak numbers.
- Storage temperature affected SELDI-TOF-MS profiles, while centrifugation and freeze-thaw cycles had minimal impact.
Conclusions:
- SELDI-TOF-MS is a rapid, high-throughput technique suitable for profiling low-mass (<10 kDa) saliva proteins/peptides.
- Pre-analytical factors critically affect saliva proteome profiling results.
- This technique provides insights into human saliva proteome composition and its temporal changes.
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