Development of a method for absolute quantification of equine acute phase proteins using concatenated peptide standards and selected reaction monitoring

Louise Bundgaard; Stine Jacobsen; Thomas Franck Dyrlund; Mette Aamand Sørensen; Victoria M. Harman; Robert J. Beynon; Philip J. Brownridge; Lars Jelstrup Petersen; Emøke Bendixen

doi:10.1021/pr500607s

Development of a method for absolute quantification of equine acute phase proteins using concatenated peptide standards and selected reaction monitoring

Louise Bundgaard, Stine Jacobsen, Thomas Franck Dyrlund, Mette Aamand Sørensen, Victoria M. Harman, Robert J. Beynon, Philip J. Brownridge, Lars Jelstrup Petersen, Emøke Bendixen

10 Citations (Scopus)

Abstract

The aim of this study was the development of a quantitative assay that could support future studies of a panel of acute phase proteins (APPs) in the horse. The assay was based on a quantification concatamer (QconCAT) coupled to selected reaction monitoring methodology. Thirty-two peptides, corresponding to 13 putative or confirmed APPs for the Equus caballus (equine) species were selected for the design of a QconCAT construct. The gene encoding the QconCAT was synthesized and expressed as an isotope-labeled chimaeric protein in Escherichia coli. The QconCAT tryptic peptides were analyzed on a triple-quadrupole instrument, and the quantotypic properties were assessed in equine serum, wound tissue, and wound interstitial fluid. Reasonable quantotypic performance was found for 12, 14, and 14 peptides in serum, wound tissue, and interstitial fluid, respectively. Seven proteins were quantified in absolute terms in serum collected from a horse before and after the onset of a systemic inflammatory condition, and the observed protein concentrations were in close agreement with previous data. We conclude, that this QconCAT is applicable for concurrent quantitative analysis of multiple APPs in serum and may also support future studies of these proteins in other types of tissues and body fluids from the horse.

Original language	English
Journal	Journal of Proteome Research
Volume	13
Issue number	12
Pages (from-to)	5635-5647
Number of pages	13
ISSN	1535-3893
DOIs	https://doi.org/10.1021/pr500607s
Publication status	Published - 5 Dec 2014

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10.1021/pr500607s

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Bundgaard, L., Jacobsen, S., Dyrlund, T. F., Sørensen, M. A., Harman, V. M., Beynon, R. J., Brownridge, P. J., Petersen, L. J., & Bendixen, E. (2014). Development of a method for absolute quantification of equine acute phase proteins using concatenated peptide standards and selected reaction monitoring. Journal of Proteome Research, 13(12), 5635-5647. https://doi.org/10.1021/pr500607s

Development of a method for absolute quantification of equine acute phase proteins using concatenated peptide standards and selected reaction monitoring. / Bundgaard, Louise ; Jacobsen, Stine; Dyrlund, Thomas Franck et al.

In: Journal of Proteome Research, Vol. 13, No. 12, 05.12.2014, p. 5635-5647.

Research output: Contribution to journal › Journal article › Research › peer-review

Bundgaard, L , Jacobsen, S, Dyrlund, TF, Sørensen, MA, Harman, VM, Beynon, RJ, Brownridge, PJ, Petersen, LJ & Bendixen, E 2014, 'Development of a method for absolute quantification of equine acute phase proteins using concatenated peptide standards and selected reaction monitoring', Journal of Proteome Research, vol. 13, no. 12, pp. 5635-5647. https://doi.org/10.1021/pr500607s

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title = "Development of a method for absolute quantification of equine acute phase proteins using concatenated peptide standards and selected reaction monitoring",

abstract = "The aim of this study was the development of a quantitative assay that could support future studies of a panel of acute phase proteins (APPs) in the horse. The assay was based on a quantification concatamer (QconCAT) coupled to selected reaction monitoring methodology. Thirty-two peptides, corresponding to 13 putative or confirmed APPs for the Equus caballus (equine) species were selected for the design of a QconCAT construct. The gene encoding the QconCAT was synthesized and expressed as an isotope-labeled chimaeric protein in Escherichia coli. The QconCAT tryptic peptides were analyzed on a triple-quadrupole instrument, and the quantotypic properties were assessed in equine serum, wound tissue, and wound interstitial fluid. Reasonable quantotypic performance was found for 12, 14, and 14 peptides in serum, wound tissue, and interstitial fluid, respectively. Seven proteins were quantified in absolute terms in serum collected from a horse before and after the onset of a systemic inflammatory condition, and the observed protein concentrations were in close agreement with previous data. We conclude, that this QconCAT is applicable for concurrent quantitative analysis of multiple APPs in serum and may also support future studies of these proteins in other types of tissues and body fluids from the horse.",

author = "Louise Bundgaard and Stine Jacobsen and Dyrlund, {Thomas Franck} and S{\o}rensen, {Mette Aamand} and Harman, {Victoria M.} and Beynon, {Robert J.} and Brownridge, {Philip J.} and Petersen, {Lars Jelstrup} and Em{\o}ke Bendixen",

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T1 - Development of a method for absolute quantification of equine acute phase proteins using concatenated peptide standards and selected reaction monitoring

AU - Bundgaard, Louise

AU - Jacobsen, Stine

AU - Dyrlund, Thomas Franck

AU - Sørensen, Mette Aamand

AU - Harman, Victoria M.

AU - Beynon, Robert J.

AU - Brownridge, Philip J.

AU - Petersen, Lars Jelstrup

AU - Bendixen, Emøke

PY - 2014/12/5

Y1 - 2014/12/5

N2 - The aim of this study was the development of a quantitative assay that could support future studies of a panel of acute phase proteins (APPs) in the horse. The assay was based on a quantification concatamer (QconCAT) coupled to selected reaction monitoring methodology. Thirty-two peptides, corresponding to 13 putative or confirmed APPs for the Equus caballus (equine) species were selected for the design of a QconCAT construct. The gene encoding the QconCAT was synthesized and expressed as an isotope-labeled chimaeric protein in Escherichia coli. The QconCAT tryptic peptides were analyzed on a triple-quadrupole instrument, and the quantotypic properties were assessed in equine serum, wound tissue, and wound interstitial fluid. Reasonable quantotypic performance was found for 12, 14, and 14 peptides in serum, wound tissue, and interstitial fluid, respectively. Seven proteins were quantified in absolute terms in serum collected from a horse before and after the onset of a systemic inflammatory condition, and the observed protein concentrations were in close agreement with previous data. We conclude, that this QconCAT is applicable for concurrent quantitative analysis of multiple APPs in serum and may also support future studies of these proteins in other types of tissues and body fluids from the horse.

AB - The aim of this study was the development of a quantitative assay that could support future studies of a panel of acute phase proteins (APPs) in the horse. The assay was based on a quantification concatamer (QconCAT) coupled to selected reaction monitoring methodology. Thirty-two peptides, corresponding to 13 putative or confirmed APPs for the Equus caballus (equine) species were selected for the design of a QconCAT construct. The gene encoding the QconCAT was synthesized and expressed as an isotope-labeled chimaeric protein in Escherichia coli. The QconCAT tryptic peptides were analyzed on a triple-quadrupole instrument, and the quantotypic properties were assessed in equine serum, wound tissue, and wound interstitial fluid. Reasonable quantotypic performance was found for 12, 14, and 14 peptides in serum, wound tissue, and interstitial fluid, respectively. Seven proteins were quantified in absolute terms in serum collected from a horse before and after the onset of a systemic inflammatory condition, and the observed protein concentrations were in close agreement with previous data. We conclude, that this QconCAT is applicable for concurrent quantitative analysis of multiple APPs in serum and may also support future studies of these proteins in other types of tissues and body fluids from the horse.

U2 - 10.1021/pr500607s

DO - 10.1021/pr500607s

M3 - Journal article

SN - 1535-3893

VL - 13

SP - 5635

EP - 5647

JO - Journal of Proteome Research

JF - Journal of Proteome Research

IS - 12

ER -

Development of a method for absolute quantification of equine acute phase proteins using concatenated peptide standards and selected reaction monitoring

Abstract

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