Development and validation of a liquid chromatography tandem mass spectrometry assay for the quantitation of a protein therapeutic in cynomolgus monkey serum

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Yue ZhaoJim X Shen

Abstract

We have developed and fully validated a fast and simple LC-MS/MS assay to quantitate a therapeutic protein BMS-A in cynomolgus monkey serum. Prior to trypsin digestion, a recently reported sample pretreatment method was applied to remove more than 95% of the total serum albumin and denature the proteins in the serum sample. The pretreatment procedure simplified the biological sample prior to digestion, improved digestion efficiency and reproducibility, and did not require reduction and alkylation. The denatured proteins were then digested with trypsin at 60 °C for 30 min and the tryptic peptides were chromatographically separated on an Acquity CSH column (2.1 mm × 50 mm, 1.7 μm) using gradient elution. One surrogate peptide was used for quantitation and another surrogate peptide was selected for confirmation. Two corresponding stable isotope labeled peptides were used to compensate variations during LC-MS detection. The linear analytical range of the assay was 0.50-500 μg/mL. The accuracy (%Dev) was within ± 5.4% and the total assay variation (%CV) was less than 12.0% for sample analysis. The validated method demonstrated good accuracy and precision and the application of the innovative albumin removal sample pretreatment metho...Continue Reading

References

Dec 13, 2006·Mass Spectrometry Reviews·Constantin Tamvakopoulos
Nov 19, 2010·Bioanalysis·Allena Ji JiNaidong Weng
Mar 11, 2011·Bioanalysis·Eric Ezan
Jul 17, 2012·Epidemiology and Psychiatric Sciences·I Bighelli, C Barbui

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