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2020-05-12

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cc-by-nc-nd (c) Elsevier B.V., 2020
Si us plau utilitzeu sempre aquest identificador per citar o enllaçar aquest document: https://hdl.handle.net/2445/162038

Enrichment of histidine containing peptides by on-line immobilized metal affinity solid-phase extraction capillary electrophoresis-mass spectrometry

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https://doi.org/10.1016/j.microc.2020.105013

Resum

The analysis and detection of targeted peptides has received due attention in many fields of proteomics research, such as discovery of biomarkers. In this regard, capillary electrophoresis-mass spectrometry (CE-MS) is widely used, however, direct analysis of low-abundance peptides such as histidine-containing peptides (His-peptides) in complex matrixes by CE-MS remains an analytical challenge. In the present study, an immobilised metal affinity solid-phase extraction containing Ni(II) coupled on-line to capillary electrophoresis-mass spectrometry (IMA-SPE-CE-MS) method has been developed to selectively enrich His-peptides from protein tryptic digests and enhance sensitivity. The method was optimised with α-casein and validated with other standard proteins (β-casein and κ-casein). Later, it was applied to an Escherichia Coli (E. Coli) whole cell lysate. IMA-SPE-CE-MS was very selective and allowed an enrichment factor up to 100-fold. The on-line enrichment and separation method coupled to MS detection is straightforward and advantageous over off-line pretreatment methods in terms of simplicity, cost-effectiveness and throughput.

Matèries

Electroforesi capil·lar, Espectrometria de masses, Pèptids

Matèries (anglès)

Capillary electrophoresis, Mass spectrometry, Peptides

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Articles publicats en revistes (Enginyeria Química i Química Analítica)

Citació

PERÓ GASCÓN, Roger, GIMÉNEZ LÓPEZ, Estela, SANZ NEBOT, María victoria, BENAVENTE MORENO, Fernando j. (julián). Enrichment of histidine containing peptides by on-line immobilized metal affinity solid-phase extraction capillary electrophoresis-mass spectrometry. _Microchemical Journal_. 2020. Vol. 267, núm. 121881. [consulta: 24 de gener de 2026]. ISSN: 0026-265X. [Disponible a: https://hdl.handle.net/2445/162038]

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