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

Peter Bults; Marcel Meints; Anders Sonesson; Magnus Knutsson; Rainer Bischoff; Nico C van de Merbel

doi:10.4155/bio-2018-0032

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

Peter Bults, Marcel Meints, Anders Sonesson, Magnus Knutsson, Rainer Bischoff, Nico C van de Merbel

Research output: Contribution to journal › Article › Academic › peer-review

12 Citations (Scopus)

117 Downloads (Pure)

Abstract

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 language	English
Pages (from-to)	1009-1021
Journal	Bioanalysis
Volume	10
Issue number	13
Early online date	28-Jun-2018
DOIs	https://doi.org/10.4155/bio-2018-0032
Publication status	Published - 18-Jul-2018

Access to Document

10.4155/bio-2018-0032

Improving selectivity and sensitivity of protein quantitation by LC–HR–MS / MS: determination of somatropin in rat plasmaFinal publisher's version, 1.71 MBLicence: Taverne

Handle.net

Persistent link

Cite this

@article{7c087384c2ac43da8422b6d458356917,

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

abstract = "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.",

author = "Peter Bults and Marcel Meints and Anders Sonesson and Magnus Knutsson and Rainer Bischoff and {van de Merbel}, {Nico C}",

year = "2018",

month = jul,

day = "18",

doi = "10.4155/bio-2018-0032",

language = "English",

volume = "10",

pages = "1009--1021",

journal = "Bioanalysis",

issn = "1757-6180",

publisher = "FUTURE SCI LTD",

number = "13",

}

TY - JOUR

T1 - Improving selectivity and sensitivity of protein quantitation by LC-HR-MS/MS

T2 - Determination of somatropin in rat plasma

AU - Bults, Peter

AU - Meints, Marcel

AU - Sonesson, Anders

AU - Knutsson, Magnus

AU - Bischoff, Rainer

AU - van de Merbel, Nico C

PY - 2018/7/18

Y1 - 2018/7/18

N2 - 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.

AB - 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.

U2 - 10.4155/bio-2018-0032

DO - 10.4155/bio-2018-0032

M3 - Article

C2 - 29952231

SN - 1757-6180

VL - 10

SP - 1009

EP - 1021

JO - Bioanalysis

JF - Bioanalysis

IS - 13

ER -

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

Abstract

Access to Document

Handle.net

Fingerprint

Cite this