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Ultrasensitive Quantification of Recombinant Proteins Using AAA-MS.

Mathilde Louwagie1, Sylvie Kieffer-Jaquinod1, Virginie Brun2

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|July 27, 2019
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Summary

Quantifying purified recombinant proteins is crucial for various industries. This study presents a novel method using microwave-assisted hydrolysis and high-resolution mass spectrometry for accurate protein measurement.

Keywords:
Amino acid analysisMass spectrometryProteinQuantification

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Protein Science

Background:

  • Recombinant proteins are vital in therapeutics, biotechnology, and food industries.
  • Accurate quantification of purified recombinant proteins is essential for quality control and product development.
  • Existing quantification methods may have limitations in precision or scope.

Purpose of the Study:

  • To describe a protocol for the precise quantification of purified recombinant proteins.
  • To introduce a method combining microwave-assisted acidic hydrolysis with high-resolution mass spectrometry (HRMS).
  • To enable absolute quantification using stable isotope-labeled amino acids as internal standards.

Main Methods:

  • Microwave-assisted acidic hydrolysis to break down the target protein into peptides.
  • High-resolution mass spectrometry (HRMS) for sensitive and accurate detection of hydrolytic products.
  • Absolute quantification achieved by incorporating controlled amounts of labeled amino acids as calibration standards.

Main Results:

  • The described protocol enables accurate and precise determination of recombinant protein quantity and/or concentration.
  • The combination of microwave-assisted hydrolysis and HRMS offers a robust analytical approach.
  • The use of labeled amino acid standards ensures reliable absolute quantification.

Conclusions:

  • This protocol provides a reliable method for the absolute quantification of purified recombinant proteins.
  • The technique is applicable to various recombinant proteins used in diverse industrial applications.
  • This approach enhances the quality control and characterization of biopharmaceutical and biotechnological products.