Preparative protein concentration from biofluids by epitachophoresis

Helena, Hrušková; Voráčová, Ivona; Laštovičková, Markéta; Killinger, Michael; Foret, František

doi:10.1016/j.chroma.2022.463591

Cited by 4 publications

(6 citation statements)

References 19 publications

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“…ETP již byla popsána v několika našich studiích [4][5][6][7] . Jedná se o preparativní separační metodu založenou na diskontinuální elektroforéze umožňující koncentrovat a přečistit roztoky biomakromolekul z velkých objemů (až desítky ml) biologických matric (viz obr.…”

Section: Koncentrování Biomakromolekul Pomocí Epitachoforézyunclassified

“…V práci 7 , která byla součástí soutěže o Cenu Metrohm 2023, jsme se věnovali využití ETP k zakoncentrování proteinů. Náboj proteinů je určen pH roztoku, ve kterém jsou rozpuštěny.…”

Section: Epitachoforéza Proteinůunclassified

“…Koncentrační faktor v tomto případě činil 32. V poslední fázi výzkumu se nám podařilo koncentrací živného kultivačního média HeLa buněk detegovat cytochrom c. Bez ETP předúpravy tato analýza nebyla možná 7 .…”

Section: Epitachoforéza Proteinůunclassified

“…Naše dříve dosažené výsledky zveřejněné v pracích [4][5][6][7] jednoznačně dokazují, že ETP by mohla být důležitou technikou pro získávání proteinů a DNA z biologických matric. Umožňuje podstatné zakoncentrování cílových biomolekul a zároveň je schopná převést vzorek do čistší-ho roztoku.…”

Section: Závěrunclassified

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Epitachophoresis – A Method for the Extraction of Biomacromolecules from Complex Biological Matrices

Hrušková,

Voráčková,

Laštovičková

et al. 2023

Chem. Listy

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Epitachophoresis is a new separation preparative technique based on discontinuous electrolyte systems enabling the preconcentration of biomacromolecules from complex biological matrices. Moreover, it enables the transfer of these compounds from biological liquids, such as urine or growth medium, to a simpler solution, suitable for subsequent analyses. In our recently published papers, we focused on the use of epitachophoresis to pretreat the samples containing proteins and DNA. The technique enabled more than seventyfold preconcentration with recoveries reaching more than 90%. The preparative nature of epitachophoresis enables one to subsequently analyse the isolated biomacromolecules by various techniques including mass spectrometry, capillary electrophoresis, gel electrophoresis, ELISA analysis, or new-generation sequencing. Full text English translation is available in the on-line version.

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Section: Koncentrování Biomakromolekul Pomocí Epitachoforézyunclassified

Section: Epitachoforéza Proteinůunclassified

Section: Závěrunclassified

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Epitachophoresis – A Method for the Extraction of Biomacromolecules from Complex Biological Matrices

Hrušková,

Voráčková,

Laštovičková

et al. 2023

Chem. Listy

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“…For that reason, the isolation of ctDNA fragments can enhance the performance of the subsequent PCR. 5,6 In our previous studies [7][8][9][10][11] , electromigration separation techniques proved to be a powerful tool for the extraction of biomacromolecules from complex biological matrices. Other examples of previously published systems for DNA (or other biomacromolecules) fragmentation and purification such as a preparative system from BIO-RAD laboratories (Hercules, USA), GELFREE 8100 Fractionation System (Expedeon Ltd, Cambridge, UK), commercially available cutters for DNA excision (NIPPON Genetics, Tokyo, Japan), and purification kits from Thermo Fischer Scientific (Waltham, USA), can be also mentioned.…”

Section: Introductionmentioning

confidence: 99%

3D-printed device for DNA fragmentation prior liquid biopsy testing

Hrušková,

Řemínek,

Minárik

et al. 2023

Preprint

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Direct detection of DNA in complex biological samples relates to several problems regarding the selectivity and sensitivity of analyses. Therefore, DNA pre-extraction from biofluids is an emerging tool in biologically related fields. Specifically, a newly developing family of liquid biopsy techniques using PCR detection of circulating tumor DNA from blood serum or blood plasma could be significantly improved by the harnessing of fast and high-throughput DNA sample preparation. For these reasons, we have developed a 3D-printed device and a method based on gel electrophoresis for the preparative separation of DNA fragments from blood. The introduced system successfully eliminated large DNA fragments from the samples. Recovery for short DNA fragments was reaching up to 80%. The developed method was tested on complex biological matrices as blood plasma and blood serum spiked with DNA standard, and it significantly alleviated the signal of matrix DNA.

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