BUDILOVÁ, Jana, Jiří PAZOUREK, Pavel KRÁSENSKÝ and Jan POSPÍCHAL. A CONTINUOUS MODE OF OPERATION FOR LARGE VOLUME DOSING IN ANALYTICAL CARRIER AMPHOLYTE–FREE ISOELECTRIC FOCUSING OF PROTEINS APPLIED TO OFF–LINE DETECTION OF FRACTIONS. Journal of Separation Science. Weinheim: WILEY-VCH Verlag GmbH & Co., 2006, vol. 29, No 11, p. 1613-1621. ISSN 1615-9306.
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Basic information
Original name A CONTINUOUS MODE OF OPERATION FOR LARGE VOLUME DOSING IN ANALYTICAL CARRIER AMPHOLYTE–FREE ISOELECTRIC FOCUSING OF PROTEINS APPLIED TO OFF–LINE DETECTION OF FRACTIONS
Name in Czech A CONTINUOUS MODE OF OPERATION FOR LARGE VOLUME DOSING IN ANALYTICAL CARRIER AMPHOLYTE–FREE ISOELECTRIC FOCUSING OF PROTEINS APPLIED TO OFF–LINE DETECTION OF FRACTIONS
Authors BUDILOVÁ, Jana (203 Czech Republic), Jiří PAZOUREK (203 Czech Republic, guarantor), Pavel KRÁSENSKÝ (203 Czech Republic) and Jan POSPÍCHAL (203 Czech Republic).
Edition Journal of Separation Science, Weinheim, WILEY-VCH Verlag GmbH & Co. 2006, 1615-9306.
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: 2.535
RIV identification code RIV/00216224:14310/06:00016304
Organization unit Faculty of Science
UT WoS 000239574100011
Keywords in English capillary isoelectric focusing;neutralization reaction boundary;proteins;fractionation;injected large volume sample
Tags capillary isoelectric focusing, fractionation, injected large volume sample, neutralization reaction boundary, proteins
Tags Reviewed
Changed by Changed by: doc. RNDr. Bc. Jiří Pazourek, Ph.D., učo 2356. Changed: 26/1/2007 12:55.
Abstract
Mass spectrometry (MS) is being increasingly used for analysis of proteome complex samples. To remove matrix interferences and to concentrate analytes prior to a MS measurement, sample preparation is often necessary. A useful method for this purpose is Carrier Ampholyte Free-Isoelectric Focusing (CAF-IEF). In this paper CAF-IEF of ampholytes was performed on a commercial apparatus EA101 (Villa Labeco, Slovakia) equipped with a specially made column for samples of large volumes (up to 0.5 ml). A new continuous mode without voltage interruption or electrolyte replacement was developed. In this mode, a low molecular pI marker (PIM 7.4), low concentrations of myoglobin and insulin (16 mg/l), respectively, were concentrated, and then 5-ul fractions collected for off-line analyses. The total time of focusing was 66 minutes. Concentration of PIM 7.4 in the fractions was increased up to 75 times (determined by UV-VIS spectrometry). Concentration in fractions was increased up to 30 times for myoglobin and 10 times for insulin, respectively.
Abstract (in Czech)
Mass spectrometry (MS) is being increasingly used for analysis of proteome complex samples. To remove matrix interferences and to concentrate analytes prior to a MS measurement, sample preparation is often necessary. A useful method for this purpose is Carrier Ampholyte Free-Isoelectric Focusing (CAF-IEF). In this paper CAF-IEF of ampholytes was performed on a commercial apparatus EA101 (Villa Labeco, Slovakia) equipped with a specially made column for samples of large volumes (up to 0.5 ml). A new continuous mode without voltage interruption or electrolyte replacement was developed. In this mode, a low molecular pI marker (PIM 7.4), low concentrations of myoglobin and insulin (16 mg/l), respectively, were concentrated, and then 5-ul fractions collected for off-line analyses. The total time of focusing was 66 minutes. Concentration of PIM 7.4 in the fractions was increased up to 75 times (determined by UV-VIS spectrometry). Concentration in fractions was increased up to 30 times for myoglobin and 10 times for insulin, respectively.
Links
GA305/03/1100, research and development projectName: Syntéza a studium neuroprotektivních peptidů odvozených od humaninu
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