Effect of stationary phase surface chemistry and particle
architecture in proteomics

J 2025

Effect of stationary phase surface chemistry and particle architecture in proteomics

VALÁŠEK, Jan; Lukáš HEKERLE; Martina NECHVÁTALOVÁ; Antonín BEDNAŘÍK; Jan PREISLER et al.

Základní údaje

Originální název

Effect of stationary phase surface chemistry and particle architecture in proteomics

Autoři

VALÁŠEK, Jan ; Lukáš HEKERLE; Martina NECHVÁTALOVÁ; Antonín BEDNAŘÍK; Jan PREISLER a Jiří URBAN

Vydání

Journal of Chromatography, Elsevier, 2025, 0021-9673

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10406 Analytical chemistry

Stát vydavatele

Nizozemské království

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 4.000 v roce 2024

Označené pro přenos do RIV

Ano

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

Gradient elution; Column characterization; Reversed-phase retention; Bottom-up; Proteomics; Peptide

Štítky

14313010, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 17. 12. 2025 14:30, Mgr. Pavla Foltynová, Ph.D.

Anotace

V originále

The kinetic properties of four columns packed with fully porous particles and three with superficially porous particles were characterized for possible application in proteomic bottom-up analyses. All columns provided an attachment of hydrophobic C18 chains at the surface of the stationary phase. However, they differed in the additional attachment of polar groups and/or endcapping procedure. We have used the retention modeling protocol to explore the separation efficiency and maximal achievable peak capacity on tested columns. Almost all columns provided comparable maximal peak capacity in the range of 500 - 700 for the eight-hour gradient run. This confirms that the family of the stationary phases used in the bottom-up proteomics can be extended. In the case of fully porous particles, we found that the higher the column peak capacity, the higher the number of identified peptides in the simple proteomic sample, with approximately one identified peptide per peak capacity unit. On the contrary, in the case of the superficially porous particles, the number of identified peptides in the sample decreased with the higher column peak capacity. This trend can be overturned only when the lower amount of the sample is injected. Hence, when bottom-up proteomics is considered, the lower loadability of the superficially porous particles still needs to be addressed. Most stationary phases tested can be successfully used in the bottom-up analyses. However, the stationary phases with incorporated polar functional groups reduced the undesirable contribution of free silanol groups to peptide peak tailing and increased the information provided by LC-MS analysis.

Návaznosti

GA23-07581S, projekt VaV

Název: Vícerozměrná proteomická analýza proteinů a peptidů

Investor: Grantová agentura ČR, Vícerozměrná proteomická analýza proteinů a peptidů

Citovat

VALÁŠEK, Jan; Lukáš HEKERLE; Martina NECHVÁTALOVÁ; Antonín BEDNAŘÍK; Jan PREISLER a Jiří URBAN. Effect of stationary phase surface chemistry and particle architecture in proteomics. Journal of Chromatography. Elsevier, 2025, roč. 1752, July, s. 465976-465983. ISSN 0021-9673. Dostupné z: https://doi.org/10.1016/j.chroma.2025.465976.

@article{2497657,
   author = {Valášek, Jan and Hekerle, Lukáš and Nechvátalová, Martina and Bednařík, Antonín and Preisler, Jan and Urban, Jiří},
   article_number = {July},
   doi = {https://doi.org/10.1016/j.chroma.2025.465976},
   keywords = {Gradient elution; Column characterization; Reversed-phase retention; Bottom-up; Proteomics; Peptide},
   language = {eng},
   issn = {0021-9673},
   journal = {Journal of Chromatography},
   title = {Effect of stationary phase surface chemistry and particle architecture in proteomics},
   url = {https://doi.org/10.1016/j.chroma.2025.465976},
   volume = {1752},
   year = {2025}
}

TY  - JOUR
ID  - 2497657
AU  - Valášek, Jan - Hekerle, Lukáš - Nechvátalová, Martina - Bednařík, Antonín - Preisler, Jan - Urban, Jiří
PY  - 2025
TI  - Effect of stationary phase surface chemistry and particle architecture in proteomics
JF  - Journal of Chromatography
VL  - 1752
IS  - July
SP  - 465976
EP  - 465976
PB  - Elsevier
SN  - 00219673
KW  - Gradient elution
KW  - Column characterization
KW  - Reversed-phase retention
KW  - Bottom-up
KW  - Proteomics
KW  - Peptide
UR  - https://doi.org/10.1016/j.chroma.2025.465976
N2  - The kinetic properties of four columns packed with fully porous particles and three with superficially porous particles were characterized for possible application in proteomic bottom-up analyses. All columns provided an attachment of hydrophobic C18 chains at the surface of the stationary phase. However, they differed in the additional attachment of polar groups and/or endcapping procedure. We have used the retention modeling protocol to explore the separation efficiency and maximal achievable peak capacity on tested columns. Almost all columns provided comparable maximal peak capacity in the range of 500 - 700 for the eight-hour gradient run. This confirms that the family of the stationary phases used in the bottom-up proteomics can be extended. In the case of fully porous particles, we found that the higher the column peak capacity, the higher the number of identified peptides in the simple proteomic sample, with approximately one identified peptide per peak capacity unit. On the contrary, in the case of the superficially porous particles, the number of identified peptides in the sample decreased with the higher column peak capacity. This trend can be overturned only when the lower amount of the sample is injected. Hence, when bottom-up proteomics is considered, the lower loadability of the superficially porous particles still needs to be addressed. Most stationary phases tested can be successfully used in the bottom-up analyses. However, the stationary phases with incorporated polar functional groups reduced the undesirable contribution of free silanol groups to peptide peak tailing and increased the information provided by LC-MS analysis.
ER  -

VALÁŠEK, Jan; Lukáš HEKERLE; Martina NECHVÁTALOVÁ; Antonín BEDNAŘÍK; Jan PREISLER a Jiří URBAN. Effect of stationary phase surface chemistry and particle architecture in proteomics. \textit{Journal of Chromatography}. Elsevier, 2025, roč.~1752, July, s.~465976-465983. ISSN~0021-9673. Dostupné z: https://doi.org/10.1016/j.chroma.2025.465976.

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