Preparative isoelectric focusing of microorganisms in
cellulose-based separation medium and subsequent analysis by
CIEF and MALDI-TOF MS

J 2017

Preparative isoelectric focusing of microorganisms in cellulose-based separation medium and subsequent analysis by CIEF and MALDI-TOF MS

HORKA, M., K. SLAIS, J. SALPLACHTA and Filip RŮŽIČKA

Basic information

Original name

Preparative isoelectric focusing of microorganisms in cellulose-based separation medium and subsequent analysis by CIEF and MALDI-TOF MS

Authors

HORKA, M. (203 Czech Republic), K. SLAIS (203 Czech Republic), J. SALPLACHTA (203 Czech Republic) and Filip RŮŽIČKA (203 Czech Republic, guarantor, belonging to the institution)

Edition

Analytica Chimica Acta, Amsterdam, Elsevier Science publishers, 2017, 0003-2670

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10406 Analytical chemistry

Country of publisher

Netherlands

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 5.123

RIV identification code

RIV/00216224:14110/17:00094480

Organization unit

Faculty of Medicine

DOI

http://dx.doi.org/10.1016/j.aca.2017.08.046

UT WoS

000412671400017

Keywords in English

Preparative isoelectric focusing; Colored microorganisms; Isoelectric points; CIEF and MALDI-TOF MS

Abstract

V originále

Pre-separation and pre-concentration of bacteria is an important step especially when they are uncultured and bacterial concentration in the matrix is low. This study describes a preparative method based on isoelectric focusing of colored microorganisms in a cellulose-based separation medium from a high conductivity matrix. The isoelectric points found for the examined cells were 1.8 for Micrococcus luteus, 3.5 for Dietzia sp., and 4.7 for Rhodotorula mucilaginosa using capillary isoelectric focusing. The final positions of the zones of colored microbial cells in the cellulose-bed are indicated by colored pI markers. Segments of the separation medium with cells were harvested by a spatula, simply purified using centrifugation and analyzed by capillary isoelectric focusing and matrix-assisted laser desorption/ionization time of flight mass spectrometry. The determined recovery ranged from 78% to 93%. The viability of the harvested cells was verified by their cultivation.

Links

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

HORKA, M., K. SLAIS, J. SALPLACHTA and Filip RŮŽIČKA. Preparative isoelectric focusing of microorganisms in cellulose-based separation medium and subsequent analysis by CIEF and MALDI-TOF MS. Analytica Chimica Acta. Amsterdam: Elsevier Science publishers, 2017, vol. 990, OCT 16 2017, p. 185-193. ISSN 0003-2670. Available from: https://dx.doi.org/10.1016/j.aca.2017.08.046.

@article{1395637,
   author = {Horka, M. and Slais, K. and Salplachta, J. and Růžička, Filip},
   article_location = {Amsterdam},
   article_number = {OCT 16 2017},
   doi = {http://dx.doi.org/10.1016/j.aca.2017.08.046},
   keywords = {Preparative isoelectric focusing; Colored microorganisms; Isoelectric points; CIEF and MALDI-TOF MS},
   language = {eng},
   issn = {0003-2670},
   journal = {Analytica Chimica Acta},
   title = {Preparative isoelectric focusing of microorganisms in cellulose-based separation medium and subsequent analysis by CIEF and MALDI-TOF MS},
   volume = {990},
   year = {2017}
}

TY  - JOUR
ID  - 1395637
AU  - Horka, M. - Slais, K. - Salplachta, J. - Růžička, Filip
PY  - 2017
TI  - Preparative isoelectric focusing of microorganisms in cellulose-based separation medium and subsequent analysis by CIEF and MALDI-TOF MS
JF  - Analytica Chimica Acta
VL  - 990
IS  - OCT 16 2017
SP  - 185-193
EP  - 185-193
PB  - Elsevier Science publishers
SN  - 00032670
KW  - Preparative isoelectric focusing
KW  - Colored microorganisms
KW  - Isoelectric points
KW  - CIEF and MALDI-TOF MS
N2  - Pre-separation and pre-concentration of bacteria is an important step especially when they are uncultured and bacterial concentration in the matrix is low. This study describes a preparative method based on isoelectric focusing of colored microorganisms in a cellulose-based separation medium from a high conductivity matrix. The isoelectric points found for the examined cells were 1.8 for Micrococcus luteus, 3.5 for Dietzia sp., and 4.7 for Rhodotorula mucilaginosa using capillary isoelectric focusing. The final positions of the zones of colored microbial cells in the cellulose-bed are indicated by colored pI markers. Segments of the separation medium with cells were harvested by a spatula, simply purified using centrifugation and analyzed by capillary isoelectric focusing and matrix-assisted laser desorption/ionization time of flight mass spectrometry. The determined recovery ranged from 78% to 93%. The viability of the harvested cells was verified by their cultivation.
ER  -

HORKA, M., K. SLAIS, J. SALPLACHTA and Filip RŮŽIČKA. Preparative isoelectric focusing of microorganisms in cellulose-based separation medium and subsequent analysis by CIEF and MALDI-TOF MS. \textit{Analytica Chimica Acta}. Amsterdam: Elsevier Science publishers, 2017, vol.~990, OCT 16 2017, p.~185-193. ISSN~0003-2670. Available from: https://dx.doi.org/10.1016/j.aca.2017.08.046.

Detailed Information on Publication Record