Performance Assessment of a 125 Human Plasma Peptide Mixture Stored at Room Temperature for Multiple Reaction Monitoring-Mass Spectrometry

Claudia Gaither; Robert Popp; Sophia P. Borchers; Kjartan Skarphedinsson; Finnur F. Eiriksson; Margrét Thorsteinsdóttir; Yassene Mohammed; Christoph H. Borchers

doi:10.1021/acs.jproteome.1c00249

Performance Assessment of a 125 Human Plasma Peptide Mixture Stored at Room Temperature for Multiple Reaction Monitoring-Mass Spectrometry

Claudia Gaither, Robert Popp, Sophia P. Borchers, Kjartan Skarphedinsson, Finnur F. Eiriksson, Margrét Thorsteinsdóttir, Yassene Mohammed, Christoph H. Borchers^*

^*Corresponding author for this work

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

Abstract

Synthetic peptides are a critical requirement for the development and application of targeted mass spectrometry (MS)-based assays for the quantitation of proteins from biological matrices. Transporting synthetic peptides on dry ice from one laboratory to another is costly and often difficult because of country-specific import and export regulations. Therefore, in this study, we assessed the impact of leaving a lyophilized mixture consisting of 125 peptides at room temperature for up to 20 days, and we assessed the effect on the quantitative performance of multiple reaction monitoring-MS (MRM-MS) assays. The findings suggest that there are no significant differences in the MRM-MS results for the time points assessed in this study (up to 20 days). All the calibration curves and quality control (QC) samples met the acceptance criteria for precision and accuracy (raw data are available via the public MS data repository PanoramaWeb, identifier: /MRM Proteomics/2020_BAK125_RT). The number of endogenous proteins quantifiable across five plasma samples was consistently between 87 and 99 out of 125 for all time points. Moreover, the coefficients of variation (CVs) calculated for the majority of peptide concentrations across all samples and time points were <5%. In addition, a lyophilized peptide mixture was transported from Canada to Iceland without dry ice. The results showed that there was no significant difference in the quantitative performance, with the determined concentrations of most proteins in the samples falling within 30% between the analyses performed on the same three plasma samples in Iceland and those in Canada. Overall, a comparison of the results obtained in Canada and in Iceland indicated that the peptides were stable under the conditions tested and also indicated that shipping lyophilized peptide mixtures without dry ice, but in the presence of sufficient desiccant material, could be a feasible option in cases where transport difficulties may arise or dry-ice sublimation may occur.

Original language	English
Pages (from-to)	4292-4302
Number of pages	11
Journal	Journal of Proteome Research
Volume	20
Issue number	9
DOIs	https://doi.org/10.1021/acs.jproteome.1c00249
Publication status	Published - 3 Sept 2021

Bibliographical note

Funding Information:
MRM Proteomics Inc. is grateful to the University of Victoria, Genome BC Proteomics Centre, for their efforts in designing and validating the MRM assays for the 125 human surrogate peptides, as well as for performing CZE and AAA analyses. Additionally, we appreciate Coralie Geric’s help in coordinating the room-temperature incubations throughout the experiments. CHB and the University of Victoria-Genome British Columbia Proteomics Centre are grateful to Genome Canada for financial support through the Genomics Technology Platform (264PRO). CHB is also grateful for support from the Terry Fox Research Institute and the Segal McGill Chair in Molecular Oncology at McGill University (Montreal, Quebec, Canada). CHB is also grateful for support from the Warren Y. Soper Charitable Trust and the Alvin Segal Family Foundation to the Jewish General Hospital (Montreal, Quebec, Canada). The study was also supported by the MegaGrant of the Ministry of Science and Higher Education of the Russian Federation (Agreement with the Skolkovo Institute of Science and Technology No. 075-10-2019-083).

Publisher Copyright:
© 2021 The Authors. Published by American Chemical Society.

Other keywords

internal standards
mass spectrometry
multiple reaction monitoring (MRM)
peptide
proteomics
stability
targeted proteomics

Access to Document

10.1021/acs.jproteome.1c00249

Cite this

Gaither, C., Popp, R., Borchers, S. P., Skarphedinsson, K., Eiriksson, F. F., Thorsteinsdóttir, M., Mohammed, Y., & Borchers, C. H. (2021). Performance Assessment of a 125 Human Plasma Peptide Mixture Stored at Room Temperature for Multiple Reaction Monitoring-Mass Spectrometry. Journal of Proteome Research, 20(9), 4292-4302. https://doi.org/10.1021/acs.jproteome.1c00249

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title = "Performance Assessment of a 125 Human Plasma Peptide Mixture Stored at Room Temperature for Multiple Reaction Monitoring-Mass Spectrometry",

abstract = "Synthetic peptides are a critical requirement for the development and application of targeted mass spectrometry (MS)-based assays for the quantitation of proteins from biological matrices. Transporting synthetic peptides on dry ice from one laboratory to another is costly and often difficult because of country-specific import and export regulations. Therefore, in this study, we assessed the impact of leaving a lyophilized mixture consisting of 125 peptides at room temperature for up to 20 days, and we assessed the effect on the quantitative performance of multiple reaction monitoring-MS (MRM-MS) assays. The findings suggest that there are no significant differences in the MRM-MS results for the time points assessed in this study (up to 20 days). All the calibration curves and quality control (QC) samples met the acceptance criteria for precision and accuracy (raw data are available via the public MS data repository PanoramaWeb, identifier: /MRM Proteomics/2020_BAK125_RT). The number of endogenous proteins quantifiable across five plasma samples was consistently between 87 and 99 out of 125 for all time points. Moreover, the coefficients of variation (CVs) calculated for the majority of peptide concentrations across all samples and time points were <5%. In addition, a lyophilized peptide mixture was transported from Canada to Iceland without dry ice. The results showed that there was no significant difference in the quantitative performance, with the determined concentrations of most proteins in the samples falling within 30% between the analyses performed on the same three plasma samples in Iceland and those in Canada. Overall, a comparison of the results obtained in Canada and in Iceland indicated that the peptides were stable under the conditions tested and also indicated that shipping lyophilized peptide mixtures without dry ice, but in the presence of sufficient desiccant material, could be a feasible option in cases where transport difficulties may arise or dry-ice sublimation may occur.",

keywords = "internal standards, mass spectrometry, multiple reaction monitoring (MRM), peptide, proteomics, stability, targeted proteomics",

author = "Claudia Gaither and Robert Popp and Borchers, {Sophia P.} and Kjartan Skarphedinsson and Eiriksson, {Finnur F.} and Margr{\'e}t Thorsteinsd{\'o}ttir and Yassene Mohammed and Borchers, {Christoph H.}",

note = "Funding Information: MRM Proteomics Inc. is grateful to the University of Victoria, Genome BC Proteomics Centre, for their efforts in designing and validating the MRM assays for the 125 human surrogate peptides, as well as for performing CZE and AAA analyses. Additionally, we appreciate Coralie Geric{\textquoteright}s help in coordinating the room-temperature incubations throughout the experiments. CHB and the University of Victoria-Genome British Columbia Proteomics Centre are grateful to Genome Canada for financial support through the Genomics Technology Platform (264PRO). CHB is also grateful for support from the Terry Fox Research Institute and the Segal McGill Chair in Molecular Oncology at McGill University (Montreal, Quebec, Canada). CHB is also grateful for support from the Warren Y. Soper Charitable Trust and the Alvin Segal Family Foundation to the Jewish General Hospital (Montreal, Quebec, Canada). The study was also supported by the MegaGrant of the Ministry of Science and Higher Education of the Russian Federation (Agreement with the Skolkovo Institute of Science and Technology No. 075-10-2019-083). Publisher Copyright: {\textcopyright} 2021 The Authors. Published by American Chemical Society.",

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doi = "10.1021/acs.jproteome.1c00249",

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Gaither, C, Popp, R, Borchers, SP, Skarphedinsson, K, Eiriksson, FF, Thorsteinsdóttir, M, Mohammed, Y & Borchers, CH 2021, 'Performance Assessment of a 125 Human Plasma Peptide Mixture Stored at Room Temperature for Multiple Reaction Monitoring-Mass Spectrometry', Journal of Proteome Research, vol. 20, no. 9, pp. 4292-4302. https://doi.org/10.1021/acs.jproteome.1c00249

TY - JOUR

T1 - Performance Assessment of a 125 Human Plasma Peptide Mixture Stored at Room Temperature for Multiple Reaction Monitoring-Mass Spectrometry

AU - Gaither, Claudia

AU - Popp, Robert

AU - Borchers, Sophia P.

AU - Skarphedinsson, Kjartan

AU - Eiriksson, Finnur F.

AU - Thorsteinsdóttir, Margrét

AU - Mohammed, Yassene

AU - Borchers, Christoph H.

N1 - Funding Information: MRM Proteomics Inc. is grateful to the University of Victoria, Genome BC Proteomics Centre, for their efforts in designing and validating the MRM assays for the 125 human surrogate peptides, as well as for performing CZE and AAA analyses. Additionally, we appreciate Coralie Geric’s help in coordinating the room-temperature incubations throughout the experiments. CHB and the University of Victoria-Genome British Columbia Proteomics Centre are grateful to Genome Canada for financial support through the Genomics Technology Platform (264PRO). CHB is also grateful for support from the Terry Fox Research Institute and the Segal McGill Chair in Molecular Oncology at McGill University (Montreal, Quebec, Canada). CHB is also grateful for support from the Warren Y. Soper Charitable Trust and the Alvin Segal Family Foundation to the Jewish General Hospital (Montreal, Quebec, Canada). The study was also supported by the MegaGrant of the Ministry of Science and Higher Education of the Russian Federation (Agreement with the Skolkovo Institute of Science and Technology No. 075-10-2019-083). Publisher Copyright: © 2021 The Authors. Published by American Chemical Society.

PY - 2021/9/3

Y1 - 2021/9/3

N2 - Synthetic peptides are a critical requirement for the development and application of targeted mass spectrometry (MS)-based assays for the quantitation of proteins from biological matrices. Transporting synthetic peptides on dry ice from one laboratory to another is costly and often difficult because of country-specific import and export regulations. Therefore, in this study, we assessed the impact of leaving a lyophilized mixture consisting of 125 peptides at room temperature for up to 20 days, and we assessed the effect on the quantitative performance of multiple reaction monitoring-MS (MRM-MS) assays. The findings suggest that there are no significant differences in the MRM-MS results for the time points assessed in this study (up to 20 days). All the calibration curves and quality control (QC) samples met the acceptance criteria for precision and accuracy (raw data are available via the public MS data repository PanoramaWeb, identifier: /MRM Proteomics/2020_BAK125_RT). The number of endogenous proteins quantifiable across five plasma samples was consistently between 87 and 99 out of 125 for all time points. Moreover, the coefficients of variation (CVs) calculated for the majority of peptide concentrations across all samples and time points were <5%. In addition, a lyophilized peptide mixture was transported from Canada to Iceland without dry ice. The results showed that there was no significant difference in the quantitative performance, with the determined concentrations of most proteins in the samples falling within 30% between the analyses performed on the same three plasma samples in Iceland and those in Canada. Overall, a comparison of the results obtained in Canada and in Iceland indicated that the peptides were stable under the conditions tested and also indicated that shipping lyophilized peptide mixtures without dry ice, but in the presence of sufficient desiccant material, could be a feasible option in cases where transport difficulties may arise or dry-ice sublimation may occur.

AB - Synthetic peptides are a critical requirement for the development and application of targeted mass spectrometry (MS)-based assays for the quantitation of proteins from biological matrices. Transporting synthetic peptides on dry ice from one laboratory to another is costly and often difficult because of country-specific import and export regulations. Therefore, in this study, we assessed the impact of leaving a lyophilized mixture consisting of 125 peptides at room temperature for up to 20 days, and we assessed the effect on the quantitative performance of multiple reaction monitoring-MS (MRM-MS) assays. The findings suggest that there are no significant differences in the MRM-MS results for the time points assessed in this study (up to 20 days). All the calibration curves and quality control (QC) samples met the acceptance criteria for precision and accuracy (raw data are available via the public MS data repository PanoramaWeb, identifier: /MRM Proteomics/2020_BAK125_RT). The number of endogenous proteins quantifiable across five plasma samples was consistently between 87 and 99 out of 125 for all time points. Moreover, the coefficients of variation (CVs) calculated for the majority of peptide concentrations across all samples and time points were <5%. In addition, a lyophilized peptide mixture was transported from Canada to Iceland without dry ice. The results showed that there was no significant difference in the quantitative performance, with the determined concentrations of most proteins in the samples falling within 30% between the analyses performed on the same three plasma samples in Iceland and those in Canada. Overall, a comparison of the results obtained in Canada and in Iceland indicated that the peptides were stable under the conditions tested and also indicated that shipping lyophilized peptide mixtures without dry ice, but in the presence of sufficient desiccant material, could be a feasible option in cases where transport difficulties may arise or dry-ice sublimation may occur.

KW - internal standards

KW - mass spectrometry

KW - multiple reaction monitoring (MRM)

KW - peptide

KW - proteomics

KW - stability

KW - targeted proteomics

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DO - 10.1021/acs.jproteome.1c00249

M3 - Article

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SN - 1535-3893

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EP - 4302

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JF - Journal of Proteome Research

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ER -

Performance Assessment of a 125 Human Plasma Peptide Mixture Stored at Room Temperature for Multiple Reaction Monitoring-Mass Spectrometry

Abstract

Bibliographical note

Other keywords

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