Simplified 2-D CE-MS mapping: analysis of proteolytic digests.
ABSTRACT It has been demonstrated that CE-MS is a very useful hyphenated technique for proteomic studies. However, the huge amount of data stored in a single CE-MS run makes it necessary to account with procedures able to extract all the relevant information made available by CE-MS. In this work, we present a new and easy approach capable of generating a simplified 2-D map from CE-MS raw data. This new approach provides the automatic detection and characterization of the most abundant ions from the CE-MS data including their mass-to-charge (m/z) values, ion intensities and analysis times. It is demonstrated that visualization of CE-MS data in this simplified 2-D format allows: (i) an easy and simultaneous visual inspection of large datasets, (ii) an immediate perception of relevant differences in closely related samples, (iii) a rapid monitoring of data quality levels in different samples, and (iv) a fast discrimination between comigrating polypeptides and ESI-MS fragmentation ions. The strategy proposed in this work does not rely on an excellent mass accuracy for peak detection and filtering, since MS values obtained from an IT analyzer are used. Moreover, the methodology developed works directly with the CE-MS raw data, without interference by the user, giving simultaneously a simplified 2-D map and a much easier and more complete data evaluation. Besides, this procedure can easily be implemented in any CE-MS laboratory. The usefulness of this approach is validated by studying the very similar trypsin digests from bovine, rabbit and horse cytochrome c. It is demonstrated that this simplified 2-D approach allows specific markers for each species to be obtained in a fast and simple way.
- SourceAvailable from: Akiyoshi Hirayama[Show abstract] [Hide abstract]
ABSTRACT: Abstract In metabolomics, the rapid identification of quantitative differences between multiple biological samples remains a majorMetabolomics. 01/2010;
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ABSTRACT: The review presents a comprehensive survey of recent developments of high-performance capillary electromigration methods, zone electrophoresis, ITP, IEF, affinity electrophoresis, EKC and electrochromatography, and their application to analysis, preparation and physicochemical characterization of peptides. New approaches to the theoretical description and experimental investigation of electromigration properties of peptides, and to methodology of their separations, such as sample preparation, adsorption suppression, EOF control and detection, are described. New developments in particular CE and CEC modes are reported and several types of their applications to peptide analysis are presented: conventional qualitative and quantitative analysis, determination in biomatrices, monitoring of chemical and enzymatical reactions and physical changes, amino acid and sequence analysis, and peptide mapping of proteins. Few examples of micropreparative peptide separations are shown and capabilities of CE and CEC techniques to provide important physicochemical characteristics of peptides are demonstrated.Electrophoresis 11/2009; 31(1):122-46. · 3.26 Impact Factor
Article: Serum proteomics for gastric cancer.[Show abstract] [Hide abstract]
ABSTRACT: According to the World Health Organization, 800,000 cancer-related deaths are caused by gastric cancer each year globally, hence making it the second leading cause of cancer-related deaths in the world. Gastric cancer is often either asymptomatic or causing only nonspecific symptoms in its early stages. By the time the symptoms occur, the cancer has usually reached an advanced stage, which is one of the main reasons for its relatively poor prognosis. Therefore, early diagnosis and early treatment are very crucial. The differential analysis of serum protein between cancer patients and healthy controls can be performed using proteomics techniques and can hence be adopted as tumor biomarkers for the early diagnosis of cancer. So far, several serum tumor biomarkers have been identified for gastric cancer. However due to their poor specificity and sensitivity, they have proven to be insufficient for the reliable diagnosis of gastric cancer. Thus, using modern advanced proteomic techniques to find some new and reliable serum tumor biomarkers for earlier and reliable diagnosis of gastric cancer is a must. Nowadays, proteomics-based techniques, such as SELDI and HCLP, are available to discover biomarkers in gastric cancer. Numerous novel serum tumor biomarkers such as SAA, plasminogen and C9c, have been discovered through serological proteomics strategies. This review mainly focuses on the serum proteomics techniques and their application in the research of gastric cancer.Clinica chimica acta; international journal of clinical chemistry 02/2014; · 2.54 Impact Factor