A comparative study of synthetic winged peptides for absolute protein quantification

Abstract A proper internal standard choice is critical for accurate, precise, and reproducible mass spectrometry-based proteomics assays. Synthetic isotopically labeled (SIL) proteins are currently considered the gold standard. However, they are costly and challenging to obtain. An alternative appro...

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Autores principales:	Eliska Benesova, Veronika Vidova, Zdenek Spacil
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b72acab8bfcb4521b397ce9475f4939b
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spelling	oai:doaj.org-article:b72acab8bfcb4521b397ce9475f4939b2021-12-02T15:00:55ZA comparative study of synthetic winged peptides for absolute protein quantification10.1038/s41598-021-90087-92045-2322https://doaj.org/article/b72acab8bfcb4521b397ce9475f4939b2021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-90087-9https://doaj.org/toc/2045-2322Abstract A proper internal standard choice is critical for accurate, precise, and reproducible mass spectrometry-based proteomics assays. Synthetic isotopically labeled (SIL) proteins are currently considered the gold standard. However, they are costly and challenging to obtain. An alternative approach uses SIL peptides or SIL "winged" peptides extended at C- or/and N-terminus with an amino acid sequence or a tag cleaved during enzymatic proteolysis. However, a consensus on the design of a winged peptide for absolute quantification is missing. In this study, we used human serum albumin as a model system to compare the quantitative performance of reference SIL protein with four different designs of SIL winged peptides: (i) commercially available SIL peptides with a proprietary trypsin cleavable tag at C-terminus, (ii) SIL peptides extended with five amino acid residues at C-terminus, (iii) SIL peptides extended with three and (iv) with five amino acid residues at both C- and N-termini. Our results demonstrate properties of various SIL extended peptides designs, e.g., water solubility and efficiency of trypsin enzymatic cleavage with primary influence on quantitative performance. SIL winged peptides extended with three amino acids at both C- and N-termini demonstrated optimal quantitative performance, equivalent to the SIL protein.Eliska BenesovaVeronika VidovaZdenek SpacilNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Eliska Benesova Veronika Vidova Zdenek Spacil A comparative study of synthetic winged peptides for absolute protein quantification
description	Abstract A proper internal standard choice is critical for accurate, precise, and reproducible mass spectrometry-based proteomics assays. Synthetic isotopically labeled (SIL) proteins are currently considered the gold standard. However, they are costly and challenging to obtain. An alternative approach uses SIL peptides or SIL "winged" peptides extended at C- or/and N-terminus with an amino acid sequence or a tag cleaved during enzymatic proteolysis. However, a consensus on the design of a winged peptide for absolute quantification is missing. In this study, we used human serum albumin as a model system to compare the quantitative performance of reference SIL protein with four different designs of SIL winged peptides: (i) commercially available SIL peptides with a proprietary trypsin cleavable tag at C-terminus, (ii) SIL peptides extended with five amino acid residues at C-terminus, (iii) SIL peptides extended with three and (iv) with five amino acid residues at both C- and N-termini. Our results demonstrate properties of various SIL extended peptides designs, e.g., water solubility and efficiency of trypsin enzymatic cleavage with primary influence on quantitative performance. SIL winged peptides extended with three amino acids at both C- and N-termini demonstrated optimal quantitative performance, equivalent to the SIL protein.
format	article
author	Eliska Benesova Veronika Vidova Zdenek Spacil
author_facet	Eliska Benesova Veronika Vidova Zdenek Spacil
author_sort	Eliska Benesova
title	A comparative study of synthetic winged peptides for absolute protein quantification
title_short	A comparative study of synthetic winged peptides for absolute protein quantification
title_full	A comparative study of synthetic winged peptides for absolute protein quantification
title_fullStr	A comparative study of synthetic winged peptides for absolute protein quantification
title_full_unstemmed	A comparative study of synthetic winged peptides for absolute protein quantification
title_sort	comparative study of synthetic winged peptides for absolute protein quantification
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/b72acab8bfcb4521b397ce9475f4939b
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A comparative study of synthetic winged peptides for absolute protein quantification

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