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

页码：1009 / 1021

页数：13

共 32 条

[31] Strategy for the Quantitation of a Protein Conjugate via Hybrid Immunocapture-Liquid Chromatography with Sequential HRMS and SRM-Based LC-MS/MS Analyses [J].