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

被引：13
作者：
Bults, Peter [1 ,2 ]
Meints, Marcel [1 ]
Sonesson, Anders [3 ]
Knutsson, Magnus [3 ]
Bischoff, Rainer [2 ]
van de Merbel, Nico C. [1 ,2 ]
机构：
[1] PRA Hlth Sci, Bioanalyt Lab, Amerikaweg 18, NL-9407 TK Assen, Netherlands
[2] Univ Groningen, Dept Analyt Biochem, A Deusinglaan 1, NL-9700 AV Groningen, Netherlands
[3] Ferring Pharmaceut, Kay Fiskers Plads 11, DK-2300 Copenhagen, Denmark
来源：
BIOANALYSIS | 2018年 / 10卷 / 13期
关键词：
digestion; high-resolution mass spectrometry; HRMS; mass extraction window; MEW; rhGH; somatropin; HIGH-RESOLUTION MS; BIOLOGICAL MATRICES; THERAPEUTIC PROTEIN; MASS-SPECTROMETRY; TRIPLE QUADRUPOLE; GROWTH-HORMONE; LIQUID-CHROMATOGRAPHY; ENZYMATIC DIGESTION; SAMPLE PREPARATION; QUANTIFICATION;
D O I：
10.4155/bio-2018-0032
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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 mu l 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 IC-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.
引用
收藏
页码：1009 / 1021
页数：13
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