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Related Concept Videos

Matrix-Assisted Laser Desorption Ionization (MALDI)01:08

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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 spectrometry is widely employed in biological and medical research, as well as in fields like pharmacology and biochemistry.
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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.
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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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Peptide Identification Using Tandem Mass Spectrometry01:33

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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.
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Mass Spectrometry: Overview01:19

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Mass spectrometry is an analytical technique used to determine the molecular mass and molecular formula of a compound. The basic principle of mass spectrometry is to generate ions from the analyte molecule and measure these ion abundances against their molecular mass.  One common type of ionization, known as electrospray ionization or EI, bombards the analyte molecules in the gas phase with high-energy electron beams. The electron beams displace an electron from the molecule and leave...
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High-Throughput Label-Free Biochemical Assays Using Infrared Matrix-Assisted Desorption Electrospray Ionization Mass

Fan Pu1, Andrew J Radosevich1, James W Sawicki1

  • 1Drug Discovery Science and Technology, AbbVie Inc., North Chicago, Illinois 60064, United States.

Analytical Chemistry
|April 22, 2021
PubMed
Summary
This summary is machine-generated.

This study introduces a novel high-throughput mass spectrometry (MS) system using infrared matrix-assisted desorption electrospray ionization (IR-MALDESI) for rapid biochemical assays. The system enables direct analysis without chromatography, achieving high speeds for drug discovery screening.

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Area of Science:

  • Analytical Chemistry
  • Biochemistry
  • Mass Spectrometry

Background:

  • Mass spectrometry (MS) offers sensitive and specific biochemical assays without labels, but chromatography limits throughput.
  • Ion suppression from sample matrices necessitates chromatography, hindering high-throughput applications of MS.

Purpose of the Study:

  • To develop and demonstrate a novel direct analysis high-throughput MS system for biochemical assays.
  • To overcome throughput limitations in MS-based drug discovery by eliminating the need for chromatography.

Main Methods:

  • Development of a high-throughput MS system utilizing infrared matrix-assisted desorption electrospray ionization (IR-MALDESI).
  • Establishment of biochemical assays for wild-type isocitrate dehydrogenase 1 (IDH1), diacylglycerol kinase zeta (DGKζ), and p300 histone acetyltransferase (P300).
  • Proof-of-concept pilot screen of approximately 3,000 compounds for IDH1 using the developed system.

Main Results:

  • The IR-MALDESI system demonstrated a potential acquisition rate of 33 spectra/s.
  • Successful development of biochemical assays in standard buffers suitable for high-throughput lead discovery.
  • Reliable data acquisition at speeds amenable for screening large compound libraries, validated by an IDH1 pilot screen.

Conclusions:

  • The novel IR-MALDESI MS system enables high-throughput direct analysis, overcoming previous limitations.
  • This technology is suitable for a broad range of lead discovery assays, significantly advancing drug screening capabilities.
  • The system provides a viable and efficient platform for large-scale compound library screening.