Preparative continuous flow electrophoretic instrumentation for purification of biological samples

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081715%3A_____%2F21%3A00544943" target="_blank" >RIV/68081715:_____/21:00544943 - isvavai.cz</a>

Výsledek na webu

<a href="http://hdl.handle.net/11104/0321728" target="_blank" >http://hdl.handle.net/11104/0321728</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1002/elps.202100160" target="_blank" >10.1002/elps.202100160</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Preparative continuous flow electrophoretic instrumentation for purification of biological samples

Popis výsledku v původním jazyce

We constructed a preparative instrumentation and developed the methods that are based on separation of the samples by bidirectional isotachophoresis/moving boundary electrophoresis in continuous divergent flow. The described instrumentation can be used for a variety of the samples, however, it can be easily optimized and tailored for the specific sample. The trapezoid separation bed from nonwoven textile exhibited minimum adsorption effect for sample and it can be used repeatedly. By the addition of different spacers via separation space inlets, the sections of pH gradient can be modified to enhance the separation. The liquid flow from two inlets positioned on each side of the sample inlet prevented the contact of the sample with anolyte and catholyte at the analysis beginning. One pair of thin electrodes (graphite and stainless-steel) was placed at the separation space output. The electrode products were washed out into drains without disturbing the focusing process. The influence of EOF was managed by tilting the separation bed in the direction from cathodic to anodic side. The components of spirulina supernatant and color pI markers were separated in the pH gradient from 3.9 to 10.1. pH gradient was stable for at least 4.5 h and spirulina supernatant from about 0.12 g of dry powder was processed. Compared to other preparative methods used for spirulina separation, the presented method/instrumentation working with a continuous divergent flow had essential advantages. The efficient separation was fast, and no intermediate steps were necessary to obtain liquid fractions with separated components compatible with further biological experiments.

Název v anglickém jazyce

Preparative continuous flow electrophoretic instrumentation for purification of biological samples

Popis výsledku anglicky

We constructed a preparative instrumentation and developed the methods that are based on separation of the samples by bidirectional isotachophoresis/moving boundary electrophoresis in continuous divergent flow. The described instrumentation can be used for a variety of the samples, however, it can be easily optimized and tailored for the specific sample. The trapezoid separation bed from nonwoven textile exhibited minimum adsorption effect for sample and it can be used repeatedly. By the addition of different spacers via separation space inlets, the sections of pH gradient can be modified to enhance the separation. The liquid flow from two inlets positioned on each side of the sample inlet prevented the contact of the sample with anolyte and catholyte at the analysis beginning. One pair of thin electrodes (graphite and stainless-steel) was placed at the separation space output. The electrode products were washed out into drains without disturbing the focusing process. The influence of EOF was managed by tilting the separation bed in the direction from cathodic to anodic side. The components of spirulina supernatant and color pI markers were separated in the pH gradient from 3.9 to 10.1. pH gradient was stable for at least 4.5 h and spirulina supernatant from about 0.12 g of dry powder was processed. Compared to other preparative methods used for spirulina separation, the presented method/instrumentation working with a continuous divergent flow had essential advantages. The efficient separation was fast, and no intermediate steps were necessary to obtain liquid fractions with separated components compatible with further biological experiments.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10406 - Analytical chemistry

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2021

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Electrophoresis

ISSN

0173-0835

e-ISSN

1522-2683

Svazek periodika

42

Číslo periodika v rámci svazku

20

Stát vydavatele periodika

DE - Spolková republika Německo

Počet stran výsledku

9

Strana od-do

2103-2111

Kód UT WoS článku

000686392800001

EID výsledku v databázi Scopus

2-s2.0-85113178523