Continuous fast focusing in a trapezoidal void channel based on bidirectional isotachophoresis in a wide pH range

Miroslava Stastna, Karel Slais

Research output: Contribution to journalArticle

Abstract

This study concentrates on development of instrumentation for focusing and separation of analytes in continuous flow. It is based on bidirectional ITP working in wide pH range with separation space of closed void channel of trapezoidal shape and continuous supply of sample. The novel instrumentation is working with electrolyte system formulated previously and on the contrary to devices currently available, it allows preparative separation and concentration of cationic, anionic, and amphoteric analytes simultaneously and in wide pH range. The formation of sharp edges at zone boundaries as well as low conductivity zones are avoided in suggested system and thus, local overheating is eliminated allowing for high current densities at initial stages of focusing. This results in high focusing speed and reduction of analysis time, which is particularly advantageous for separations performed in continuous flow systems. The closed void channel is designed to avoid basic obstacles related to liquid leakage, bubbles formation, contacts with electrodes, channel height and complicated assembling. The performance of designed instrumentation and focusing dynamics were tested by using colored low molecular mass pH indicators for local pH determination, focusing pattern, and completion. In addition, feasibility and separation efficiency were demonstrated by focusing of cytochrome C and myoglobin. The collection of fractions at instrument output allows for subsequent analysis and identification of sample components that are concentrated and conveniently in form of solution for further processing. Since the instrumentation operates with commercially available simple defined buffers and compounds without need of carrier ampholytes background, it is economically favorable.

Original languageEnglish (US)
Pages (from-to)2579-2586
Number of pages8
JournalElectrophoresis
Volume36
Issue number20
DOIs
StatePublished - Oct 1 2015
Externally publishedYes

Fingerprint

Isotachophoresis
Ampholyte Mixtures
Inosine Triphosphate
Myoglobin
Cytochromes
Bubble formation
Electrolytes
Leakage (fluid)
Molecular mass
Buffers
Electrodes
Contacts (fluid mechanics)
Current density
Equipment and Supplies
Liquids
Processing

Keywords

  • Bidirectional isotachophoresis
  • Focusing
  • Proteins
  • Trapezoidal void channel
  • Wide pH range

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry

Cite this

Continuous fast focusing in a trapezoidal void channel based on bidirectional isotachophoresis in a wide pH range. / Stastna, Miroslava; Slais, Karel.

In: Electrophoresis, Vol. 36, No. 20, 01.10.2015, p. 2579-2586.

Research output: Contribution to journalArticle

@article{11443a0158f6434f97167f5227bce8e3,
title = "Continuous fast focusing in a trapezoidal void channel based on bidirectional isotachophoresis in a wide pH range",
abstract = "This study concentrates on development of instrumentation for focusing and separation of analytes in continuous flow. It is based on bidirectional ITP working in wide pH range with separation space of closed void channel of trapezoidal shape and continuous supply of sample. The novel instrumentation is working with electrolyte system formulated previously and on the contrary to devices currently available, it allows preparative separation and concentration of cationic, anionic, and amphoteric analytes simultaneously and in wide pH range. The formation of sharp edges at zone boundaries as well as low conductivity zones are avoided in suggested system and thus, local overheating is eliminated allowing for high current densities at initial stages of focusing. This results in high focusing speed and reduction of analysis time, which is particularly advantageous for separations performed in continuous flow systems. The closed void channel is designed to avoid basic obstacles related to liquid leakage, bubbles formation, contacts with electrodes, channel height and complicated assembling. The performance of designed instrumentation and focusing dynamics were tested by using colored low molecular mass pH indicators for local pH determination, focusing pattern, and completion. In addition, feasibility and separation efficiency were demonstrated by focusing of cytochrome C and myoglobin. The collection of fractions at instrument output allows for subsequent analysis and identification of sample components that are concentrated and conveniently in form of solution for further processing. Since the instrumentation operates with commercially available simple defined buffers and compounds without need of carrier ampholytes background, it is economically favorable.",
keywords = "Bidirectional isotachophoresis, Focusing, Proteins, Trapezoidal void channel, Wide pH range",
author = "Miroslava Stastna and Karel Slais",
year = "2015",
month = "10",
day = "1",
doi = "10.1002/elps.201500223",
language = "English (US)",
volume = "36",
pages = "2579--2586",
journal = "Electrophoresis",
issn = "0173-0835",
publisher = "Wiley-VCH Verlag",
number = "20",

}

TY - JOUR

T1 - Continuous fast focusing in a trapezoidal void channel based on bidirectional isotachophoresis in a wide pH range

AU - Stastna, Miroslava

AU - Slais, Karel

PY - 2015/10/1

Y1 - 2015/10/1

N2 - This study concentrates on development of instrumentation for focusing and separation of analytes in continuous flow. It is based on bidirectional ITP working in wide pH range with separation space of closed void channel of trapezoidal shape and continuous supply of sample. The novel instrumentation is working with electrolyte system formulated previously and on the contrary to devices currently available, it allows preparative separation and concentration of cationic, anionic, and amphoteric analytes simultaneously and in wide pH range. The formation of sharp edges at zone boundaries as well as low conductivity zones are avoided in suggested system and thus, local overheating is eliminated allowing for high current densities at initial stages of focusing. This results in high focusing speed and reduction of analysis time, which is particularly advantageous for separations performed in continuous flow systems. The closed void channel is designed to avoid basic obstacles related to liquid leakage, bubbles formation, contacts with electrodes, channel height and complicated assembling. The performance of designed instrumentation and focusing dynamics were tested by using colored low molecular mass pH indicators for local pH determination, focusing pattern, and completion. In addition, feasibility and separation efficiency were demonstrated by focusing of cytochrome C and myoglobin. The collection of fractions at instrument output allows for subsequent analysis and identification of sample components that are concentrated and conveniently in form of solution for further processing. Since the instrumentation operates with commercially available simple defined buffers and compounds without need of carrier ampholytes background, it is economically favorable.

AB - This study concentrates on development of instrumentation for focusing and separation of analytes in continuous flow. It is based on bidirectional ITP working in wide pH range with separation space of closed void channel of trapezoidal shape and continuous supply of sample. The novel instrumentation is working with electrolyte system formulated previously and on the contrary to devices currently available, it allows preparative separation and concentration of cationic, anionic, and amphoteric analytes simultaneously and in wide pH range. The formation of sharp edges at zone boundaries as well as low conductivity zones are avoided in suggested system and thus, local overheating is eliminated allowing for high current densities at initial stages of focusing. This results in high focusing speed and reduction of analysis time, which is particularly advantageous for separations performed in continuous flow systems. The closed void channel is designed to avoid basic obstacles related to liquid leakage, bubbles formation, contacts with electrodes, channel height and complicated assembling. The performance of designed instrumentation and focusing dynamics were tested by using colored low molecular mass pH indicators for local pH determination, focusing pattern, and completion. In addition, feasibility and separation efficiency were demonstrated by focusing of cytochrome C and myoglobin. The collection of fractions at instrument output allows for subsequent analysis and identification of sample components that are concentrated and conveniently in form of solution for further processing. Since the instrumentation operates with commercially available simple defined buffers and compounds without need of carrier ampholytes background, it is economically favorable.

KW - Bidirectional isotachophoresis

KW - Focusing

KW - Proteins

KW - Trapezoidal void channel

KW - Wide pH range

UR - http://www.scopus.com/inward/record.url?scp=84943659468&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84943659468&partnerID=8YFLogxK

U2 - 10.1002/elps.201500223

DO - 10.1002/elps.201500223

M3 - Article

C2 - 26104601

AN - SCOPUS:84943659468

VL - 36

SP - 2579

EP - 2586

JO - Electrophoresis

JF - Electrophoresis

SN - 0173-0835

IS - 20

ER -