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Related Experiment Video

Updated: Dec 17, 2025

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
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Affinity-Bead Assisted Mass Spectrometry (Affi-BAMS): A Multiplexed Microarray Platform for Targeted Proteomics.

Ghaith M Hamza1,2, Vladislav B Bergo3, Sergey Mamaev3

  • 1Discovery Sciences, BioPharmaceutical R&D, AstraZeneca, Boston, MA 02451, USA.

International Journal of Molecular Sciences
|March 20, 2020
PubMed
Summary
This summary is machine-generated.

A new Affinity-Bead Assisted Mass Spectrometry (Affi-BAMS) technology enables quantitative, multiplexed proteomic assays. This method combines immuno-affinity capture with MALDI MS for high-throughput analysis of proteins and post-translational modifications (PTMs).

Keywords:
BAMSMALDI MSPTMsbead assisted mass spectrometrymultiplex assaystargeted proteomics

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Area of Science:

  • Proteomics and Mass Spectrometry
  • Biochemical Assays
  • Analytical Chemistry

Background:

  • Quantitative analysis of proteins and post-translational modifications (PTMs) is crucial for biological, biochemical, and pharmacological research.
  • Existing methods may lack the throughput or multiplexing capability required for comprehensive proteomic studies.
  • There is a need for advanced analytical technologies to profile multiple analytes simultaneously across numerous samples.

Purpose of the Study:

  • To introduce and validate a novel microarray analytical technology, Affinity-Bead Assisted Mass Spectrometry (Affi-BAMS).
  • To demonstrate the capability of Affi-BAMS for highly multiplexed, targeted proteomic assays.
  • To showcase the application of Affi-BAMS in profiling multiple proteins and PTMs in response to various stimuli.

Main Methods:

  • Development of a novel workflow combining immuno-affinity capture using antibody-coated beads with Matrix Assisted Laser Desorption Ionization Mass Spectrometry (MALDI MS).
  • Utilizing single-bead enrichment for specific analyte capture, enabling quantification over three orders of magnitude.
  • Implementing a spatially localized elution method for arrayed beads, creating micro spots for direct MALDI MS analysis.
  • Focusing on bottom-up proteomics for high-throughput screening of hundreds of samples per day.

Main Results:

  • Affi-BAMS successfully enables highly multiplexed and targeted proteomic assays without liquid chromatography (LC).
  • The technology supports simultaneous profiling of multiple proteins and PTMs with high specificity and customizable assay panels.
  • Demonstrated robustness and diverse applications through studies on key signaling proteins (4EBP1, RPS6, ERK1/ERK2, mTOR, Histone H3, C-MET) under various stimuli.

Conclusions:

  • Affi-BAMS represents a significant advancement in proteomic analysis, offering high throughput and multiplexing capabilities.
  • The technology facilitates quantitative profiling of proteins and PTMs, aiding complex biological and pharmacological investigations.
  • Affi-BAMS provides a powerful tool for screening large numbers of samples, accelerating discovery in proteomics research.