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Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification
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Absolute protein quantification based on calibrated particle counting using electrospray-differential mobility

Wei Mi1, Xinyi Zhang1, Bin Wang1

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

Analytica Chimica Acta
|April 18, 2024
PubMed
Summary

Accurate protein quantification is crucial for diagnostics and drugs. This study introduces a calibrated particle counting method using electrospray-differential mobility analysis (ES-DMA) and a condensation particle counter (CPC) for precise protein measurement.

Keywords:
Detection efficiencyElectrospray-differential mobility analysis-condensation particle counterParticle countingProtein quantification

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Biophysics

Background:

  • Accurate protein quantification is essential for the traceability of in vitro diagnostics and drug products.
  • Existing methods may face challenges with diverse protein sizes and lack of reference standards.

Purpose of the Study:

  • To develop an absolute quantification approach for proteins using calibrated particle counting.
  • To validate the method's performance with certified reference materials and various protein types.

Main Methods:

  • Utilized electrospray-differential mobility analysis (ES-DMA) coupled with a condensation particle counter (CPC) for calibrated particle counting.
  • Determined detection efficiency using calibrators and verified performance with Bovine Serum Albumin (BSA) and NIMCmAb reference materials.
  • Investigated the linear relationship between detection efficiency and particle size distribution for different proteins.

Main Results:

  • The ES-DMA-CPC method enabled absolute quantification of proteins based on measured particle numbers and determined detection efficiencies.
  • Measurement performance was validated using certified reference materials (BSA, NIMCmAb).
  • A linear relationship was observed between protein detection efficiency and particle size, applicable to various proteins including myoglobin, fibrinogen, NIST reference material 8671, D-dimer, and glutathione S-transferase dimer.

Conclusions:

  • The calibrated particle counting method provides a rapid, accurate, and nondestructive approach for protein quantification.
  • This method is particularly effective for measuring proteinaceous particles of varying sizes, even without specific reference standards.