Related Experiment Video
Updated: Aug 7, 2026
09:04
Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification
Published on: August 17, 2015
Surface plasmon resonance mass spectrometry for protein analysis
Dobrin Nedelkov1, Randall W Nelson
1Intrinsic Bioprobes Inc., Tempe, AZ, USA.
Methods in Molecular Biology (Clifton, N.J.)
|June 21, 2006
Summary
Surface Plasmon Resonance (SPR) and Mass Spectrometry (MS) together offer a powerful method for protein interaction analysis. This combined SPR-MS technique reveals quantitative, kinetic, and structural protein details, enhancing understanding of biological processes.
Area of Science:
- Biochemistry and Biophysics
- Analytical Chemistry
- Proteomics
Background:
- Surface Plasmon Resonance (SPR) is a label-free optical sensing technique used to study biomolecular interactions in real-time.
- Mass Spectrometry (MS) provides high-resolution mass measurements, enabling detailed structural characterization of proteins and their modifications.
- Integrating SPR and MS offers complementary information, overcoming limitations of each individual technique for comprehensive protein analysis.
Purpose of the Study:
- To present detailed procedures and protocols for performing combined Surface Plasmon Resonance and Mass Spectrometry (SPR-MS) analyses.
- To illustrate the practical application of SPR-MS methodology using cardiac troponin I (cTnI) as a model system.
- To highlight the advantages of SPR-MS for investigating protein interactions and structural integrity.
Main Methods:
- Surface Plasmon Resonance (SPR) for real-time monitoring of binding kinetics and quantitative variations.
- Mass Spectrometry (MS) for structural elucidation, including post-translational modifications and protein identification.
- Sequential or hyphenated SPR-MS workflows for comprehensive analysis of protein-ligand interactions.
Main Results:
- Demonstrated successful application of SPR-MS for analyzing cardiac troponin I (cTnI) interactions.
- Identified quantitative and kinetic binding parameters using SPR.
- Characterized structural features and potential modifications of cTnI using MS, complementing SPR data.
Conclusions:
- The combined SPR-MS approach provides a robust and versatile platform for in-depth protein interaction studies.
- This methodology allows for the simultaneous assessment of binding kinetics, quantification, and structural details, offering a more complete picture than either technique alone.
- SPR-MS is particularly valuable for detecting subtle protein modifications that may not be apparent through SPR analysis alone.
Related Concept Videos
01:19MALDI-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...
01:33Proteomics
A proteome is the entire set of proteins that a cell type produces. We can study proteomes using the knowledge of genomes because genes code for mRNAs, and the mRNAs encode proteins. Although mRNA analysis is a step in the right direction, not all mRNAs are translated into proteins.
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term proteomics...
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term proteomics...
01:33Peptide 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...
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...