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@article{1138793, author = {Duša, Filip and Šlais, Karel}, article_location = {Hoboken}, article_number = {11}, doi = {http://dx.doi.org/10.1002/elps.201200485}, keywords = {Bioactive peptides; Caseinomacropeptide; Micropreparative; Solution isoelectric focusing; Whey}, language = {eng}, issn = {0173-0835}, journal = {Electrophoresis}, title = {New solution IEF device for micropreparative separation of peptides and proteins}, url = {http://onlinelibrary.wiley.com/doi/10.1002/elps.201200485/pdf}, volume = {34}, year = {2013} }
TY - JOUR ID - 1138793 AU - Duša, Filip - Šlais, Karel PY - 2013 TI - New solution IEF device for micropreparative separation of peptides and proteins JF - Electrophoresis VL - 34 IS - 11 SP - 1519-1525 EP - 1519-1525 PB - WILEY-BLACKWELL SN - 01730835 KW - Bioactive peptides KW - Caseinomacropeptide KW - Micropreparative KW - Solution isoelectric focusing KW - Whey UR - http://onlinelibrary.wiley.com/doi/10.1002/elps.201200485/pdf L2 - http://www.ncbi.nlm.nih.gov/pubmed/23417644 N2 - The article presents a new concept of preparative solution IEF where time requirements and efficiency are similar to gel-based IEF whereas simple fraction handling as well as quick and complete protein recovery typical for solution-based IEF methods are maintained. The presented method is based on the IEF in separation medium soaked in a segmented strip of nonwoven fabric. The strip is positioned in an open horizontal V-shaped trough. Suggested focusing method combines free solution IEF under continuous evaporation and whole channel dispensing. Separation medium based on ethylene glycol/water mixture enhances viscosity enough to reduce electroosmosis and prevents the medium from completely drying out. Generation of pH gradient and final local pH is visually traced by colored low-molecular pI markers added to input mixture, which enables an optimization of focusing process and collection of individual fractions at desired pH range. The proposed method was tested by fractionation of the proteins and bioactive peptides originating from raw whey. Moreover, subsequent HPLC analysis of the individually collected solution IEF fractions was used for identification of whey components. We confirmed that the method is capable to process directly few tenths of milliliters of raw samples including the salty ones. ER -
DUŠA, Filip and Karel ŠLAIS. New solution IEF device for micropreparative separation of peptides and proteins. \textit{Electrophoresis}. Hoboken: WILEY-BLACKWELL, 2013, vol.~34, No~11, p.~1519-1525. ISSN~0173-0835. Available from: https://dx.doi.org/10.1002/elps.201200485.
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