Improving selectivity and sensitivity of protein quantitation by LC-HR-MS/MS: determination of somatropin in rat plasma
| Autor: | Anders Sonesson, Peter Bults, Nico C van de Merbel, Marcel Meints, Rainer Bischoff, Magnus Knutsson |
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| Přispěvatelé: | Analytical Biochemistry, Medicinal Chemistry and Bioanalysis (MCB) |
| Rok vydání: | 2018 |
| Předmět: |
Clinical Biochemistry
Quantitative proteomics Peptide 030226 pharmacology & pharmacy 01 natural sciences Analytical Chemistry Matrix (chemical analysis) 03 medical and health sciences 0302 clinical medicine High complexity Tandem Mass Spectrometry Animals General Pharmacology Toxicology and Pharmaceutics chemistry.chemical_classification Chromatography Human Growth Hormone 010401 analytical chemistry General Medicine 0104 chemical sciences Rats Medical Laboratory Technology Somatropin chemistry Selectivity Digestion Chromatography Liquid |
| Zdroj: | Bioanalysis, 10(13), 1009-1021. FUTURE SCI LTD |
| ISSN: | 1757-6199 1757-6180 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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