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Related Concept Videos

MALDI-TOF Mass Spectrometry01:19

MALDI-TOF Mass Spectrometry

Mass spectrometry is a powerful characterization technique that can identify and separate a wide variety of compounds ranging from chemical to biological entities, based on their mass-to-charge ratio (m/z). The instruments that allow this detection, known as mass spectrometers, have three components: an ion source, a mass analyzer, and a detector. These spectrometers differ based on the nature of their ion source and analyzers.Matrix-assisted laser desorption ionization (MALDI) is a commonly...
Peptide Identification Using Tandem Mass Spectrometry01:33

Peptide Identification Using Tandem Mass Spectrometry

Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
Electrospray Ionization (ESI) Mass Spectrometry01:12

Electrospray Ionization (ESI) Mass Spectrometry

Higher molecular weight biomolecules are nonvolatile compounds that may decompose before ionizing or vaporizing during mass analysis with conventional electron impact ionization methods. Accordingly, electrospray ionization (ESI) is the favored method for vaporizing and ionizing biomolecules as it circumvents rapid fragmentation and enables the recording of mass signals for the entire biomolecule.
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High-Resolution Mass Spectrometry (HRMS)01:15

High-Resolution Mass Spectrometry (HRMS)

The resolution of a mass spectrometer depends on the efficiency of separating ions with different ion masses. The mass of an atom is approximated to the sum of the masses of protons and neutrons inside, considering the masses of protons and neutrons as equal. However, the masses of the proton (1.6726 × 10−24 g) and neutron (1.6749 × 10−24 g) are not truly equal. There is a minor error in the expression of atomic masses relative to the simplest atom of hydrogen. For example, the mass of helium...
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Rapid Identification of Pathogens

MALDI-TOF MS has transformed clinical microbiology by offering a rapid and reliable method for pathogen identification. The traditional approach to microbial identification typically involves time-consuming culture techniques and biochemical tests, which can delay the initiation of appropriate antimicrobial therapy. MALDI-TOF MS avoids these delays by using characteristic ribosomal protein mass patterns of microbial cells, enabling accurate species-level identification within minutes.Principle...
Matrix-Assisted Laser Desorption Ionization (MALDI)01:08

Matrix-Assisted Laser Desorption Ionization (MALDI)

Matrix-assisted laser desorption ionization (MALDI) is a powerful analytical technique used in mass spectrometry. It enables the identification and characterization of various biomolecules, including proteins, peptides, nucleic acids, and carbohydrates. MALDI is an ionization technique, widely employed in biological and medical research, as well as in fields like pharmacology and biochemistry.The analyte of interest, a biomolecule or a mixture of biomolecules, is mixed with a suitable matrix...

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Dithranol as a Matrix for Matrix Assisted Laser Desorption/Ionization Imaging on a Fourier Transform Ion Cyclotron Resonance Mass Spectrometer
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Protein identification using nano-HPLC-MS: ESI-MS and MALDI-MS interfaces.

Rui Vitorino1, Jana Krenkova, Frantisek Foret

  • 1Department of Chemistry, Mass Spectrometry Center, QOPNA, University of Aveiro, Campus Universitário de Santiago, Aveiro, Portugal. rvitorino@ua.pt

Methods in Molecular Biology (Clifton, N.J.)
|September 28, 2011
PubMed
Summary

New proteomics protocols effectively characterize high molecular weight peptides and intact proteins from body fluids and tissues. These methods improve protein identification and analysis using on-line digestion and mass spectrometry.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Proteomics

Background:

  • Traditional proteomics methods struggle to characterize high molecular weight peptides and intact proteins in biological samples.
  • Complex biological matrices like body fluids and tissues present challenges for comprehensive protein analysis.

Purpose of the Study:

  • To develop and present two novel protocols for the characterization of high molecular weight peptides (>3 kDa) and intact proteins.
  • To overcome limitations of conventional proteomics techniques in analyzing complex biological samples.

Main Methods:

  • On-line trypsin digestion followed by nano-high-performance liquid chromatography (nano-HPLC) separation.
  • Mass spectrometry analysis utilizing matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI).
  • Protein denaturation in an aqueous-organic solvent to reduce sample derivatization and enhance analysis.

Main Results:

  • Successful characterization of high molecular weight peptides and intact proteins previously difficult to analyze.
  • Demonstrated reduction in sample derivatization, enabling more in-depth protein detection and identification.
  • Integration of protocols into standard proteomic workflows with high-resolution monolithic columns.

Conclusions:

  • The described protocols offer an advanced approach for comprehensive proteomic analysis of complex biological samples.
  • These methods facilitate the detection and identification of challenging high molecular weight proteins and peptides.
  • The protocols are advantageous for analyzing minimal sample amounts and integrating into existing workflows.