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Related Concept Videos

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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Two-dimensional Gel Electrophoresis01:22

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Two-dimensional gel electrophoresis is a high-resolution protein separation method first introduced by O' Farrell and Klose in 1975. This method involves protein separation by two dimensions, mass and charge, making it more accurate than one-dimensional gel electrophoresis.
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such...
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Electrophoresis: Overview01:20

Electrophoresis: Overview

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Electrophoresis is a powerful analytical separation technique that relies on the differential migration of charged species when subjected to an electric field. The core strength of electrophoresis lies in its ability to separate high-molecular-weight species in complex mixtures. It has found widespread use in biochemistry, molecular biology, and analytical chemistry, allowing the separation of compounds like amino acids, nucleotides, carbohydrates, and proteins with excellent resolution.
There...
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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.
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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Related Experiment Video

Updated: Jul 20, 2025

An HS-MRM Assay for the Quantification of Host-cell Proteins in Protein Biopharmaceuticals by Liquid Chromatography Ion Mobility QTOF Mass Spectrometry
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An HS-MRM Assay for the Quantification of Host-cell Proteins in Protein Biopharmaceuticals by Liquid Chromatography Ion Mobility QTOF Mass Spectrometry

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Development of a potential primary method for protein quantification via electrospray differential mobility analysis.

Wei Mi1, Xinyi Zhang1, Xiangrong Tian2

  • 1National Institute of Metrology, No.18 Beisanhuan Donglu, Beijing, 100029, China.

Talanta
|August 4, 2023
PubMed
Summary

A new protein quantification method using electrospray-differential mobility analysis coupled with a condensation particle counter (ES-DMA-CPC) offers accurate measurements. This ES-DMA-CPC method shows promise as a primary quantification technique, complementing existing isotope dilution mass spectrometry (IDMS) approaches.

Keywords:
Electrospray-differential mobility analysis-condensation particle counter (ES-DMA-CPC)Isotope dilution mass spectrometry (IDMS)Potential primary methodProtein quantificationUV‒VIS spectrophotometry

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Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry
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Area of Science:

  • Metrology
  • Analytical Chemistry
  • Biophysics

Background:

  • Accurate protein quantification is crucial for in vitro diagnostics and drug development.
  • Establishing metrological traceability ensures reliability in biological measurements.
  • Existing methods may have limitations, particularly for proteins of varying molecular weights.

Purpose of the Study:

  • To establish and validate a novel primary method for protein quantification.
  • To assess the analytical performance and measurement uncertainty of the proposed method.
  • To compare the new method with established techniques like isotope dilution mass spectrometry (IDMS).

Main Methods:

  • Development of a protein quantification method based on electrospray-differential mobility analysis coupled with a condensation particle counter (ES-DMA-CPC).
  • Validation using certified reference material (NIMCmAb) and evaluation of measurement uncertainty.
  • Application to diverse proteins including myoglobin, bovine serum albumin, IgG monoclonal antibody, and fibrinogen (17–340 kDa).

Main Results:

  • The ES-DMA-CPC method demonstrated accurate protein quantification across a wide molecular weight range.
  • The method showed good agreement with the gold-standard isotope dilution mass spectrometry (IDMS) method.
  • Measurement uncertainty was evaluated, confirming the reliability of the quantification.

Conclusions:

  • The ES-DMA-CPC method is a viable potential primary method for protein quantification.
  • This technique offers accurate measurements, especially for large molecular weight proteins.
  • The ES-DMA-CPC method can serve as a valuable complement to IDMS for protein analysis.