Electrophoretic techniques for purification, separation and
detection of Kayvirus with subsequent control by
matrix-assisted laser desorption/ionization time-of-flight mass
spectrometry and microbiological methods

J 2018

Electrophoretic techniques for purification, separation and detection of Kayvirus with subsequent control by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and microbiological methods

HORKÁ, Marie, Dana ŠTVERÁKOVÁ, Jiří ŠALPLACHTA, Karel ŠLAIS, Marta ŠIBOROVÁ et. al.

Basic information

Original name

Electrophoretic techniques for purification, separation and detection of Kayvirus with subsequent control by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and microbiological methods

Authors

HORKÁ, Marie (203 Czech Republic, guarantor), Dana ŠTVERÁKOVÁ (203 Czech Republic, belonging to the institution), Jiří ŠALPLACHTA (203 Czech Republic), Karel ŠLAIS (203 Czech Republic), Marta ŠIBOROVÁ (203 Czech Republic, belonging to the institution), Filip RŮŽIČKA (203 Czech Republic, belonging to the institution) and Roman PANTŮČEK (203 Czech Republic, belonging to the institution)

Edition

Journal of Chromatography A, Amsterdam, Netherlands, Elsevier, 2018, 0021-9673

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10609 Biochemical research methods

Country of publisher

Netherlands

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 3.858

RIV identification code

RIV/00216224:14310/18:00105970

Organization unit

Faculty of Science

DOI

http://dx.doi.org/10.1016/j.chroma.2018.07.078

UT WoS

000445982400018

Keywords in English

Capillary electrophoretic methods; Preparative IEF; Purification; Pre-concentration; Separation; Kayvirus; Staphylococcus aureus; MALDI-TOF MS

Abstract

V originále

The bacteriophage K1/420 is a member of genus Kayvirus that was extensively studied as an alternative treatment to combat bacterial infections caused by antibiotic-resistant Staphylococcus aureus strains. Despite the promise of phage therapy, the development of clinical applications of phages is facing regulatory and technical hurdles before it can receive acceptance in the Western World. Suitable simple and accurate diagnostic techniques to control the quality of the phage, which would satisfy the requirements of regulatory authorities are still being discussed. Here, we present the conditions for the simultaneous separation and detection of phage K1/420 and S. aureus by CZE and by CIEF were found, and the phage isoelectric point was determined to be 3.6. After removing the cell debris, the phage was successfully purified from the crude phage lysate and pre-concentrated by preparative isoelectric focusing. Its zone was localized by the positions of colored pI markers in the cellulose bed. The phage from the harvested zone had a decreased ability to infect its host. However, it was suitable for its separation, detection and identification by capillary electrophoretic methods, MALDI-TOF MS and electron microscopy. (C) 2018 Elsevier B.V. All rights reserved.

Links

LQ1601, research and development project

Name: CEITEC 2020 (Acronym: CEITEC2020)

Investor: Ministry of Education, Youth and Sports of the CR

NV16-29916A, research and development project

Name: Využití bakteriofágů v léčbě nozokomiálních infekcí spojených s multirezistencí či tvorbou biofilmu

VI20172020069, research and development project

Name: Vývoj moderních instrumentálních metod pro rychlou detekci a identifikaci vybraných B-agens a toxinů (Acronym: VMIMRDIBT)

Investor: Ministry of the Interior of the CR

Citovat

HORKÁ, Marie, Dana ŠTVERÁKOVÁ, Jiří ŠALPLACHTA, Karel ŠLAIS, Marta ŠIBOROVÁ, Filip RŮŽIČKA and Roman PANTŮČEK. Electrophoretic techniques for purification, separation and detection of Kayvirus with subsequent control by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and microbiological methods. Journal of Chromatography A. Amsterdam, Netherlands: Elsevier, 2018, vol. 1570, OCT, p. 155-163. ISSN 0021-9673. Available from: https://dx.doi.org/10.1016/j.chroma.2018.07.078.

@article{1491997,
   author = {Horká, Marie and Štveráková, Dana and Šalplachta, Jiří and Šlais, Karel and Šiborová, Marta and Růžička, Filip and Pantůček, Roman},
   article_location = {Amsterdam, Netherlands},
   article_number = {OCT},
   doi = {http://dx.doi.org/10.1016/j.chroma.2018.07.078},
   keywords = {Capillary electrophoretic methods; Preparative IEF; Purification; Pre-concentration; Separation; Kayvirus; Staphylococcus aureus; MALDI-TOF MS},
   language = {eng},
   issn = {0021-9673},
   journal = {Journal of Chromatography A},
   title = {Electrophoretic techniques for purification, separation and detection of Kayvirus with subsequent control by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and microbiological methods},
   url = {https://www.sciencedirect.com/science/article/pii/S0021967318309658},
   volume = {1570},
   year = {2018}
}

TY  - JOUR
ID  - 1491997
AU  - Horká, Marie - Štveráková, Dana - Šalplachta, Jiří - Šlais, Karel - Šiborová, Marta - Růžička, Filip - Pantůček, Roman
PY  - 2018
TI  - Electrophoretic techniques for purification, separation and detection of Kayvirus with subsequent control by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and microbiological methods
JF  - Journal of Chromatography A
VL  - 1570
IS  - OCT
SP  - 155-163
EP  - 155-163
PB  - Elsevier
SN  - 00219673
KW  - Capillary electrophoretic methods
KW  - Preparative IEF
KW  - Purification
KW  - Pre-concentration
KW  - Separation
KW  - Kayvirus
KW  - Staphylococcus aureus
KW  - MALDI-TOF MS
UR  - https://www.sciencedirect.com/science/article/pii/S0021967318309658
L2  - https://www.sciencedirect.com/science/article/pii/S0021967318309658
N2  - The bacteriophage K1/420 is a member of genus Kayvirus that was extensively studied as an alternative treatment to combat bacterial infections caused by antibiotic-resistant Staphylococcus aureus strains. Despite the promise of phage therapy, the development of clinical applications of phages is facing regulatory and technical hurdles before it can receive acceptance in the Western World. Suitable simple and accurate diagnostic techniques to control the quality of the phage, which would satisfy the requirements of regulatory authorities are still being discussed. Here, we present the conditions for the simultaneous separation and detection of phage K1/420 and S. aureus by CZE and by CIEF were found, and the phage isoelectric point was determined to be 3.6. After removing the cell debris, the phage was successfully purified from the crude phage lysate and pre-concentrated by preparative isoelectric focusing. Its zone was localized by the positions of colored pI markers in the cellulose bed. The phage from the harvested zone had a decreased ability to infect its host. However, it was suitable for its separation, detection and identification by capillary electrophoretic methods, MALDI-TOF MS and electron microscopy. (C) 2018 Elsevier B.V. All rights reserved.
ER  -

HORKÁ, Marie, Dana ŠTVERÁKOVÁ, Jiří ŠALPLACHTA, Karel ŠLAIS, Marta ŠIBOROVÁ, Filip RŮŽIČKA and Roman PANTŮČEK. Electrophoretic techniques for purification, separation and detection of Kayvirus with subsequent control by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and microbiological methods. \textit{Journal of Chromatography A}. Amsterdam, Netherlands: Elsevier, 2018, vol.~1570, OCT, p.~155-163. ISSN~0021-9673. Available from: https://dx.doi.org/10.1016/j.chroma.2018.07.078.

Detailed Information on Publication Record