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Applicability of analytical and preparative monolithic columns to the separation and isolation of major whey proteinsAlbreht, Alen ; Vovk, Irena, 1965-The separation and isolation of major whey proteins is already extensively covered in the literature although no study has been published in which monolithic columns were used. In our research we ... present, for the first time, the use of short convective interaction media (CIM) monolithic columns for theseparation of all major whey proteins and isolation of ?-lactoglobulin variant A and B (?-LgA and ?-LgB) from a commercial product whey isolate (WI).Although our primary interest was directed towards finding a proper monolithic column and chromatographic conditions for the purification and isolation of ?-LgA and ?-LgB, three additional analytical LC methods, each having its own potential application target, were also developed in the courseof our research. On the monolithic diethylaminoethyl convective interaction media analytical column (CIMac DEAE), the separation of major wheyproteins was achieved by gradually lowering the pH of the mobile phase. The ever-so-hard obtainable linear external pH gradient was very linear in therange of pH 5.5-3 and the developed ion-exchange (IE) high-performance liquid chromatographic (HPLC) method was amenable to mass spectrometry (MS). Avery fast baseline separation, with UV detection, of all major whey proteins was achieved on a prototype CIMac reversed-phase styrene-divinylbenzene (RP-SDVB) monolithic column in only 4 min and the performance of this column proved superior in comparison with the packed particle POROS perfusion column.The developed RP-HPLC-MS method is fast and, due to the MS detector, can offer low limits of detection and quantitation. Finally, in order to fulfill our primary interest, a scale-up method was developed, using a prototype 8 mL analogue of the CIMac RP-SDVB column, for the isolation of native and chemically unmodified ?-LgA and ?-LgB from WI with purities higher than 90% and 81%, respectively. The proteins were to be used in further protein-ligand binding studies. The developed methods excel in speed of the analysis, sensitivity, resolution, and simplicity. Thus, it is shown for the first time that short monolithic columns are applicable to the separation and isolation of major whey proteins and that their use has some obvious benefitsVir: Journal of chromatography. A. - ISSN 0021-9673 (Vol. 1227, 2012, str. 210-218)Vrsta gradiva - članek, sestavni delLeto - 2012Jezik - angleškiCOBISS.SI-ID - 4909850
Avtor
Albreht, Alen |
Vovk, Irena, 1965-
Teme
Whey proteins |
Bovine serum albumin |
HPLC |
MS |
CIM |
CIMac monolith
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Leto | Faktor vpliva | Izdaja | Kategorija | Razvrstitev | ||||
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JCR | SNIP | JCR | SNIP | JCR | SNIP | JCR | SNIP |
Baze podatkov, v katerih je revija indeksirana
Ime baze podatkov | Področje | Leto |
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Povezave do osebnih bibliografij avtorjev | Povezave do podatkov o raziskovalcih v sistemu SICRIS |
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Albreht, Alen | 29488 |
Vovk, Irena, 1965- | 11395 |
Vir: Osebne bibliografije
in: SICRIS
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