Chromatographic Techniques for the Separation of Peptides: Application to Proteomics

Gedela, Srinubabu; Medicherla, Narasimha Rao

doi:10.1365/s10337-007-0215-9

Cited by 13 publications

References 118 publications

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Separation of Peptide Diastereomers

Scriba

2014

Encyclopedia of Analytical Chemistry

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Diastereomeric peptides, isomers in which one or more of the chiral centers have been converted to the opposite configuration (R or S, D or L ), often possess different biological activities and/or conformational properties. Therefore, the separation and determination of diastereomeric peptides is important in life sciences and particularly significant to the pharmaceutical industry for quality control of peptide synthesis and stability as well as for regulatory requirements. Diastereomers differ in their physicochemical properties that can be utilized for their determination by numerous techniques. In contrast to many other methods such as optical rotation, circular dichroism, differential scanning calorimetry, or infrared (IR) spectroscopy, separation techniques such as chromatography or capillary electrophoresis (CE) allow the sensitive and simultaneous identification and quantification of peptide diastereomers. This chapter covers analytical‐scale separation of peptide diastereomers by chromatographic methods, including paper chromatography (PC), thin‐layer chromatography (TLC), gas chromatography (GC), high‐performance liquid chromatography (HPLC), and CE. HPLC is currently the predominantly used method for peptide diastereomer separations. However, owing to the high resolving power, particularly for polar compounds, CE has been applied to the separation of diastereomeric peptides in recent years. The chapter focuses on methods employing achiral stationary phases or buffers without chiral additives for the separations, although some examples of such separations, i.e. the use of chiral columns or chiral mobile phase additives, are mentioned. Moreover, only diasteromeric peptides containing natural amino acids will be discussed.

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Separation of Peptide Diastereomers

Scriba

2014

Encyclopedia of Analytical Chemistry

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Fundamental Chemical and Structural Principles

2009

Peptides: Chemistry and Biology

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Trends in proteomic analysis of human vitreous humor samples

et al. 2014

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Proteomic analysis of human vitreous humor (VH) may elucidate the pathogenesis of retinal ocular diseases and may provide information for the development of potential therapeutic targets due to its pivotal location near lens and retina. The discovery of whole VH proteome involves a complex analysis of thousands of proteins simultaneously. Therefore, in proteomic studies the protein fractionation is important for reducing sample complexity, facilitating the access to the low-abundant proteins, and recognizing them as biotargets for clinical research. Although several separation methods have been used, gel-based proteomics are the most popular and versatile ones applied for global protein separation. However, chromatographic methods and its combination with other separation techniques are now beginning to be used as promising set-ups for VH protein identification. This review attempts to offer an overview of the techniques currently used with VH, exploring its methodological demands, exposing its advantages, and helping the reader to plan future experiences. Moreover, this review shows the relevance of VH proteomic analysis as a tool for the study of the mechanisms underlying some ocular diseases and for the development of new therapeutic approaches.

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Chromatographic Techniques for the Separation of Peptides: Application to Proteomics

Cited by 13 publications

References 118 publications

Separation of Peptide Diastereomers

Separation of Peptide Diastereomers

Fundamental Chemical and Structural Principles

Trends in proteomic analysis of human vitreous humor samples

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