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Peptide Identification Using Tandem Mass Spectrometry01:33

Peptide Identification Using Tandem Mass Spectrometry

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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.
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...
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Mass Spectrometry: Overview01:19

Mass Spectrometry: Overview

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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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Tandem Mass Spectrometry01:21

Tandem Mass Spectrometry

959
Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and signal-to-noise ratio for the analyte. Instruments with multiple analyzers separated by an interaction cell enable secondary fragmentation and selected study of the fragment ions.
Secondary fragmentations occur in the interaction cell and can be induced by various factors. Fragmentation induced by collision with inert gases, such as N2, Ar, He, etc., is called collision-induced...
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MALDI-TOF Mass Spectrometry01:19

MALDI-TOF Mass Spectrometry

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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.
Matrix-assisted laser desorption ionization (MALDI) is a commonly...
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Electrospray Ionization (ESI) Mass Spectrometry01:12

Electrospray Ionization (ESI) Mass Spectrometry

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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.
ESI utilizes electrical energy to transfer ions from the liquid phase of the sample into the...
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Mass Spectrometers01:16

Mass Spectrometers

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This lesson details the instrumentation of a mass spectrometer—a physical instrument to perform mass spectrometry on analyte molecules and record the characteristic mass spectra. This is achieved via three chief functions:
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アコースティック・エジェクション・マススペクトロメトリーによる高通量共振変調器スクリーニング

Xiujuan Wen1, Chang Liu2, Kiersten Tovar1

  • 1Merck & Co., Inc., Rahway, New Jersey 07065, United States.

Journal of the American Chemical Society
|July 12, 2024
PubMed
まとめ

高解像度アコースティック・エジェクション・マススペクトロメトリー (AEMS) は,コヴァラント・モディファイヤーの高速で高通量スクリーニングを可能にします. この革新的なプラットフォームは 難しいタンパク質標的に対する 機能的な共性薬候補を 効率的に特定することで 薬剤発見を加速します

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科学分野:

  • 分析化学
  • 薬物の発見
  • 生物化学

背景:

  • 以前は薬で治療できなかったタンパク質を 標的とする有望な戦略です
  • 従来の質量スペクトロメトリ方法 (例えば,LC-MS,SPE-MS) は,速度およびサンプル準備の制限のために,高通量スクリーニングのボトルネックです.
  • 大規模な化合物ライブラリを共性結合物質の検出に より迅速で効率的なプラットフォームが必要である.

研究 の 目的:

  • 高解像度アコースティック・エジェクション・マススペクトロメトリー (AEMS) の試作品を導入し,評価する.
  • 複雑なサンプルを最小限の準備で分析するためのAEMSの実現可能性を評価する.
  • AEMSの性能を,共振変数スクリーニングの従来の質量スペクトロメトリ技術と比較する.

主な方法:

  • 高解像度アコースティック・エジェクション・マススペクトロメトリー (AEMS) システムのプロトタイプの開発.
  • およそ1万の共性変容体のライブラリを50 kDaの標的タンパク質 (ワーナー症候群ヘリケーズ) に対して1536-well形式でスクリーニング.
  • 浄化せずに高塩分バッファでのサンプルを直接分析し,その後,共性タンパク質-リガンド添加物の定量化のための自動データ分析を行う.

主要な成果:

  • AEMSシステムは 17時間で約1万の化合物のスクリーニングを完了しました
  • このスクリーニングは,0. 8のZ'因数と2. 3%のヒット率で高精度を示した.
  • 後に行われた生化学検査で,AEMSはLC-MSと比較して偽陽性/偽陰性率が低いことが確認された.

結論:

  • 試作のAEMSプラットフォームは,コヴァラント変容体の頑丈で高通量スクリーニングを可能にします.
  • AEMSは,サンプル準備を最小限に抑えながら,既存の方法と比較してスループット (10〜100倍) と図書館の容量を大幅に増加させます.
  • この革新的なアプローチは,困難な治療目標のための機能的共性薬候補の特定を加速します.