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@article{709086, author = {Budilová, Jana and Pazourek, Jiří and Krásenský, Pavel and Pospíchal, Jan}, article_location = {Weinheim}, article_number = {11}, keywords = {capillary isoelectric focusing;neutralization reaction boundary;proteins;fractionation;injected large volume sample}, language = {eng}, issn = {1615-9306}, journal = {Journal of Separation Science}, title = {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}, volume = {29}, year = {2006} }
TY - JOUR ID - 709086 AU - Budilová, Jana - Pazourek, Jiří - Krásenský, Pavel - Pospíchal, Jan PY - 2006 TI - 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 JF - Journal of Separation Science VL - 29 IS - 11 SP - 1613-1621 EP - 1613-1621 PB - WILEY-VCH Verlag GmbH & Co. SN - 16159306 KW - capillary isoelectric focusing;neutralization reaction boundary;proteins;fractionation;injected large volume sample N2 - 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. ER -
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. \textit{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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