New solution IEF device for micropreparative separation of
peptides and proteins

J 2013

New solution IEF device for micropreparative separation of peptides and proteins

DUŠA, Filip and Karel ŠLAIS

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

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10608 Biochemistry and molecular biology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

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

Abstract

V originále

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 project

Name: 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

Citovat

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.

@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.

Detailed Information on Publication Record