Membrane-Based Nanoscale Proteolytic Reactor Enabling Protein Digestion, Peptide Separation, and Protein Identification Using Mass Spectrometry

2003 ◽  
Vol 75 (5) ◽  
pp. 1067-1074 ◽  
Author(s):  
Jonathan W. Cooper ◽  
Jinzhi Chen ◽  
Yan Li ◽  
Cheng S. Lee
Keyword(s):  
Mass Spectrometry ◽  
Protein Identification ◽  
Protein Digestion ◽  
Peptide Separation

scholarly journals Vibratory Reaction Unit for the Rapid Analysis of Proteins and Glycochains

2007 ◽  
Vol 2 ◽  
pp. 117739010700200 ◽  
Author(s):  
Yukie Sasakura ◽  
Makoto Nogami ◽  
Noriko Kobayashi ◽  
Katsuhiro Kanda
Keyword(s):  
Mass Spectrometry ◽  
Solid Surface ◽  
Protein Identification ◽  
Immobilized Enzymes ◽  
Rapid Analysis ◽  
Single Step ◽  
Protein Digestion ◽  
Sequence Coverage ◽  
Micro Scale ◽  
Reaction Unit

A protein digestion system using immobilized enzymes for protein identification and glycochain analyses has been developed, and a vibration reaction unit for micro-scale sample convection on an enzyme-immobilized solid surface was constructed. BSA as a model substrate was digested by this unit, and was successfully identified by mass spectrometry (MS) analyses. Compared to the conventional liquid-phase digestion, the reaction unit increased the number of matched peptides from 9 to 26, protein score from 455 to 1247, and sequence coverage from 21% to 48%. Glycopeptidase F (NGF), an enzyme that cleaves N-glycans from glycoproteins, was also immobilized and used to remove the glycochains from human immunoglobulin G (IgG). Trypsin and NGF were immobilized on the same solid surface and used to remove glycochains from IgG in single-step. Glycochains were labeled with fluorescent reagent and analyzed by HPLC. Several peaks corresponding to the glycochains of IgG were detected. These results suggested that the single-step digestion system, by immobilized multiple enzymes (trypsin and NGF) would be effective for the rapid structural analysis of glycoproteins.

Multiwell in-gel protein digestion and microscale sample preparation for protein identification by mass spectrometry

PROTEOMICS ◽  
2002 ◽  
Vol 2 (2) ◽  
pp. 145-150 ◽  
Author(s):  
Malcolm G. Pluskal ◽  
Alla Bogdanova ◽  
Mary Lopez ◽  
Sara Gutierrez ◽  
Aldo M. Pitt
Keyword(s):  
Mass Spectrometry ◽  
Sample Preparation ◽  
Protein Identification ◽  
Protein Digestion

scholarly journals A Review: Protein Identification by LC-MS: Principles, Instrumentation, and Applications

2020 ◽  
pp. 2448-2466
Author(s):  
Abeer Salh Alhendi
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
High Performance ◽  
Protein Identification ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Peptide Separation ◽  
Efficient Separation ◽  
Fundamental Understanding

This review will focus on protein and peptide separation studies of the period 1995 to 2010. Peptide and protein analysis have developed dramatically after applying mass spectrometry (MS) technology and other related techniques, such as two-dimensional liquid chromatography and two-dimensional gel electrophoresis. Mass spectrometry involves measurements of mass-to-charge ratios of the ionized sample. High-performance liquid chromatography (HPLC) is an important technique that is usually applied before MS is conducted due to its efficient separation. Characterization of proteins provides a foundation for the fundamental understanding of biology aspects. In this review, instrumentation, principle, applications, developments, and accuracy of the measurements of mass spectrometry will be reviewed and discussed. In addition, the principles of HPLC technology will be explained, which is necessary before applying MS.

Integrated Microfluidic System Enabling Protein Digestion, Peptide Separation, and Protein Identification

2001 ◽  
Vol 73 (11) ◽  
pp. 2648-2655 ◽  
Author(s):  
Jun Gao ◽  
Jingdong Xu ◽  
Laurie E. Locascio ◽  
Cheng S. Lee
Keyword(s):  
Protein Identification ◽  
Microfluidic System ◽  
Protein Digestion ◽  
Peptide Separation

Mass Spectrometry Techniques: Principles and Practices for Quantitative Proteomics

2020 ◽  
Vol 21 ◽  
Author(s):  
Rocco J. Rotello ◽  
Timothy D. Veenstra
Keyword(s):  
Mass Spectrometry ◽  
Quantitative Proteomics ◽  
Protein Identification ◽  
Dynamic Nature ◽  
Complete Coverage ◽  
The Past ◽  
Proteome Level ◽  
A Cell ◽  
Quantitative Changes ◽  
Made In

: In the current omics-age of research, major developments have been made in technologies that attempt to survey the entire repertoire of genes, transcripts, proteins, and metabolites present within a cell. While genomics has led to a dramatic increase in our understanding of such things as disease morphology and how organisms respond to medications, it is critical to obtain information at the proteome level since proteins carry out most of the functions within the cell. The primary tool for obtaining proteome-wide information on proteins within the cell is mass spectrometry (MS). While it has historically been associated with the protein identification, developments over the past couple of decades have made MS a robust technology for protein quantitation as well. Identifying quantitative changes in proteomes is complicated by its dynamic nature and the inability of any technique to guarantee complete coverage of every protein within a proteome sample. Fortunately, the combined development of sample preparation and MS methods have made it capable to quantitatively compare many thousands of proteins obtained from cells and organisms.

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