DUŠA, Filip and Karel ŠLAIS. New solution IEF device for micropreparative separation of peptides and proteins. 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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Basic information
Original name New solution IEF device for micropreparative separation of peptides and proteins
Authors DUŠA, Filip (203 Czech Republic, belonging to the institution) and Karel ŠLAIS (203 Czech Republic).
Edition Electrophoresis, Hoboken, WILEY-BLACKWELL, 2013, 0173-0835.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 10608 Biochemistry and molecular biology
Country of publisher United States of America
Confidentiality degree is not subject to a state or trade secret
WWW Full Text Full Text
Impact factor Impact factor: 3.161
RIV identification code RIV/00216224:14310/13:00080516
Organization unit Faculty of Science
Doi http://dx.doi.org/10.1002/elps.201200485
UT WoS 000319939300009
Keywords in English Bioactive peptides; Caseinomacropeptide; Micropreparative; Solution isoelectric focusing; Whey
Tags AKR, rivok
Tags International impact, Reviewed
Changed by Changed by: Mgr. Marie Šípková, DiS., učo 437722. Changed: 18/12/2019 13:13.
Abstract
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.
Links
VG20112015021, research and development projectName: Vývoj instrumentálních metodických postupů rychlé detekce a identifikace biologických agens v reálných vzorcích (Acronym: Detekce a identifikace biol. agens)
Investor: Ministry of the Interior of the CR
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