Journal ArticleDOI
Immobilized trypsin systems coupled on-line to separation methods: recent developments and analytical applications.
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TLDR
This review reports applications of immobilized trypsin reactors in which the IMER has been integrated into separation systems such as reversed-phase liquid chromatography or capillary electrophoresis, prior to MS analysis.Abstract:
The ability to rapidly and efficiently digest and identify an unknown protein is of great utility for proteome studies. Identification of proteins via peptide mapping is generally accomplished through proteolytic digestion with enzymes such as trypsin. Limitations of this approach consist in manual sample manipulation steps and extended reaction times for proteolytic digestion. The use of immobilized trypsin for cleavage of proteins is advantageous in comparison with application of its soluble form. Enzymes can be immobilized on different supports and used in flow systems such as immobilized enzyme reactors (IMERs). This review reports applications of immobilized trypsin reactors in which the IMER has been integrated into separation systems such as reversed-phase liquid chromatography or capillary electrophoresis, prior to MS analysis. Immobilization procedures including supports, mode of integration into separation systems, and methods are described.read more
Citations
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Journal ArticleDOI
Enzyme immobilization: an update
TL;DR: This paper is a review of the recent literatures on enzyme immobilization by various techniques, the need for immobilization and different applications in industry, covering the last two decades.
Journal ArticleDOI
Enzyme Immobilization: An Overview on Nanoparticles as ImmobilizationMatrix
Razi Ahmad,Meryam Sardar +1 more
TL;DR: This review presents the current scenario and techniques in enzyme immobilization, and the main methods used to combine proteins/enzymes with nanoparticles is given in the study.
Journal ArticleDOI
Protein digestion: an overview of the available techniques and recent developments.
TL;DR: An overview is given of the currently available digestion strategies and recent developments in the acceleration of the digestion process, and tailored approaches for classes of proteins that pose specific challenges are discussed.
Journal ArticleDOI
Effect of the support and experimental conditions in the intensity of the multipoint covalent attachment of proteins on glyoxyl-agarose supports: Correlation between enzyme–support linkages and thermal stability
Justo Pedroche,María del Mar Yust,Cesar Mateo,Roberto Fernandez-Lafuente,Julio Girón-Calle,Manuel Alaiz,Javier Vioque,Jose M. Guisan,Francisco Millán +8 more
TL;DR: It was observed that the number of lysine residues that took part in the immobilization process was a consequence of the type of support and reaction time of the experimental conditions, and that the increasing of the thermal stability of the derivatives was correlated with a increasing number ofLysines residues involved in a multipoint covalent attachment.
Journal ArticleDOI
Organic-inorganic hybrid silica monolith based immobilized trypsin reactor with high enzymatic activity.
TL;DR: A novel kind of immobilized trypsin reactor based on organic-inorganic hybrid silica monoliths was developed and successfully applied to the digestion of a mixture of model proteins and proteins extracted from E. coli.
References
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Journal ArticleDOI
An Automated Multidimensional Protein Identification Technology for Shotgun Proteomics
TL;DR: An automated method for shotgun proteomics named MudPIT, which combines multidimensional liquid chromatography with electrospray ionization tandem mass spectrometry, improves the overall analysis of proteomes by identifying proteins of all functional and physical classes.
Journal ArticleDOI
Evaluation of multidimensional chromatography coupled with tandem mass spectrometry (LC/LC-MS/MS) for large-scale protein analysis: the yeast proteome.
TL;DR: The combination of strong cation exchange (SCX) and reversed-phase (RP) chromatography to achieve two-dimensional separation prior to MS/MS and 1,504 yeast proteins were unambiguously identified in this single analysis.
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Mass Spectrometry in Proteomics
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TL;DR: This work presents strategies for Obtaining Partial Amino Acid Sequence Data from Small Quantities (5nmol) of Pure of Partially Purified Protein and Mass Spectrometric Strategies for the Structural Characterization of Proteins.
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