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@article{719268,
author = {Horká, Marie and Růžička, Filip and Holá, Veronika and Šlais, Karel},
article_location = {Heidelberg},
article_number = {5},
keywords = {capillary isoelectric focusing; microorganisms; Staphylococcus; Candida; Escherichia coli; UV detection.},
language = {eng},
issn = {1618-2642},
journal = {Analytical and Bioanalytical Chemistry},
title = {Capillary isoelectric focusing of microorganisms in the pH range 2-5 in a dynamically modified FS capillary with UV detection.},
volume = {385},
year = {2006}
}
TY - JOUR
ID - 719268
AU - Horká, Marie - Růžička, Filip - Holá, Veronika - Šlais, Karel
PY - 2006
TI - Capillary isoelectric focusing of microorganisms in the pH range 2-5 in a dynamically modified FS capillary with UV detection.
JF - Analytical and Bioanalytical Chemistry
VL - 385
IS - 5
SP - 840-846
EP - 840-846
PB - Springer-Verlag GmbH
SN - 16182642
KW - capillary isoelectric focusing
KW - microorganisms
KW - Staphylococcus
KW - Candida
KW - Escherichia coli
KW - UV detection.
N2 - The isoelectric points of many microbial cells lie within the pH range spanning from 1.5 to 4.5. In this work, we suggest a CIEF method for the separation of cells according to their isoelectric points in the pH range of 2-5. It includes the segmental injection of the sample pulse composed of the segment of the selected simple ampholytes, the segment of the bioanalytes and the segment of carrier ampholytes into fused silica capillaries dynamically modified by poly(ethylene glycole). This polymer dissolved in the catholyte, in the anolyte and in the injected sample pulse was used for a prevention of the bioanalyte adsorption on the capillary surface and for the reduction of the electroosmotic flow. Between each focusing run, the capillaries were washed with the mixture of acetone/ethanol to achieve the reproducible and efficient CIEF. In order to trace of pH gradients, low-molecular-mass pI markers were used. The mixed cultures of microorganisms, Escherichia coli CCM 3954, Candida albicans CCM 8180, Candida parapsilosis, Candida krusei, Candida glabrata, Candida tropicalis, CCM 8223, Proteus vulgaris, Klebsiela pneumoniae, Staphylococcus aureus CCM 3953, Streptococcus agalactiae CCM 6187, Enterococcus faecalis CCM 4224 and Staphylococcus epidermidis CCM 4418, were focused and separated by the CIEF method suggested here. This CIEF method enables the separation and detection of the microbes from the mixed cultures within several minutes. The minimum detectable number of microbial cells was less than 10(3).
ER -
HORKÁ, Marie, Filip RŮŽIČKA, Veronika HOLÁ and Karel ŠLAIS. Capillary isoelectric focusing of microorganisms in the pH range 2-5 in a dynamically modified FS capillary with UV detection. \textit{Analytical and Bioanalytical Chemistry}. Heidelberg: Springer-Verlag GmbH, 2006, vol.~385, No~5, p.~840-846. ISSN~1618-2642.
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