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HORKÁ, Marie, Filip RŮŽIČKA, Veronika HOLÁ and Karel ŠLAIS. CE separation of proteins and yeasts dynamically modified by PEG pyrenebutanoate with fluorescence detection. Electrophoresis. Germany: WILEY-VCH Verlag GmbH&Co. KGaA, 2007, vol. 28, No 13, p. 2300-2307, 2307 pp. ISSN 0173-0835.
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Basic information
Original name CE separation of proteins and yeasts dynamically modified by PEG pyrenebutanoate with fluorescence detection.
Name in Czech CE separace proteinů a kvasinek, dynamicky modifikovaných PEG pyrenebutanoátem, s fluorescenční detekcí
Authors HORKÁ, Marie (203 Czech Republic), Filip RŮŽIČKA (203 Czech Republic, guarantor), Veronika HOLÁ (203 Czech Republic) and Karel ŠLAIS (203 Czech Republic).
Edition Electrophoresis, Germany, WILEY-VCH Verlag GmbH&Co. KGaA, 2007, 0173-0835.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 10406 Analytical chemistry
Country of publisher Germany
Confidentiality degree is not subject to a state or trade secret
Impact factor Impact factor: 3.609
RIV identification code RIV/00216224:14110/07:00024145
Organization unit Faculty of Medicine
UT WoS 000248190400022
Keywords in English capillary isoelectric focusing; capillary electrophoresis; separation; proteins; yeasts; pyrenebutanoate
Tags Capillary electrophoresis, capillary isoelectric focusing, proteins, pyrenebutanoate, separation, yeasts
Tags International impact, Reviewed
Changed by Changed by: prof. MUDr. Filip Růžička, Ph.D., učo 1100. Changed: 25/6/2009 10:13.
Abstract
The optimized protocols of the bioanalytes separation, proteins and yeasts, dynamically modified by the nonionogenic tenside PEG pyrenebutanoate, were applied in CZE and CIEF with the acidic gradient in pH range 2-5.5, both with fluorescence detection. PEG pyrenebutanoate was used as a buffer additive for a dynamic modification of proteins and/or yeast samples. The narrow peaks of modified analytes were detected. The values of the pI's of the labeled proteins were calculated using new fluorescent pI markers in CIEF and they were found to be comparable with pI's of the native compounds. As an example of the possible use of the suggested CIEF technique, the mixed cultures of yeasts, Candida albicans, Candida glabrata, Candida kefyr, Candida krusei, Candida lusitaniae, Candida parapsilosis, Candida tropicalis, Candida zeylanoides, Geotrichum candidum, Saccharomyces cerevisiae, Trichosporon asahii and Yarrowia lipolytica, were reproducibly focused and separated with high sensitivity. Using UV excitation for the on-column fluorometric detection, the minimum detectable amounts of analytes, femtograms of proteins and down to ten cells injected on the separation capillary, were estimated.
Abstract (in Czech)
Optimalizované protokoly separace bioanalytů, proteinů a kvasinek, dynamicky upravených neionogením tenziden pyren-máselnou kyselinou, byly použity při CZE a CIEF s kyselým gradientem v pH oblasti 2 - 5.5. Použití těchto technik bylo s úspěchem ověřeno na separaci a identifikaci jednotlivých mikrobů ze směsných kultur kvasinek (Candida albicans, Candida glabrata, Candida kefyr, Candida krusei, Candida lusitaniae, Candida parapsilosis, Candida tropicalis, Candida zeylanoides, Geotrichum candidum, Saccharomyces cerevisiae, Trichosporon asahii and Yarrowia lipolytica). Používání UV excitace pro on-column fluorimetrickou detekci umožňuje detekovat jednotlivé kvasinkové buňky i minimální množství analytu (femtogramy).
Links
IAAX00310701, research and development projectName: Rychlá detekce a identifikace patogenních mikroorganizmů a virů pomocí elektromigračních technik a hmotnostní spektrometrie
Investor: Academy of Sciences of the Czech Republic
Displayed: 7/9/2024 05:28