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Improving selectivity and sensitivity of protein quantitation by LC-HR-MS/MS: Determination of somatropin in rat plasma

Bults, P., Meints, M., Sonesson, A., Knutsson, M., Bischoff, R. & van de Merbel, N. C., 18-Jul-2018, In : Bioanalysis. 10, 13, p. 1009-1021

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  • Improving selectivity and sensitivity of protein quantitation by LC–HR–MS / MS: determination of somatropin in rat plasma

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DOI

AIM: Protein quantitation by digestion of a biological sample followed by LC-MS analysis of a signature peptide can be a challenge because of the high complexity of the digested matrix. Results/methodology: The use of LC with high-resolution (quadrupole-TOF) MS detection allowed quantitation of the 22-kDa biopharmaceutical somatropin in 60 μl of rat plasma down to 25 ng/ml with minimal further sample treatment. Reducing the mass extraction window to 0.01 Da considerably decreased the interference of tryptic peptides, enhanced sensitivity and improved accuracy and precision. Analysis with LC-MS/MS resulted in a less favorable limit of quantitation of 100 ng/ml.

CONCLUSION: HRMS is an interesting option for the quantitation of proteins after digestion and has the potential to improve sensitivity with minimal method development.

Original languageEnglish
Pages (from-to)1009-1021
JournalBioanalysis
Volume10
Issue number13
Early online date28-Jun-2018
Publication statusPublished - 18-Jul-2018

ID: 62308232