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Divergent flow isoelectric focusing of whey caseinomacropeptide

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DUŠA Filip MORAVCOVÁ Dana KAHLE Vladislav ŠLAIS Karel

Rok publikování 2011
Druh Konferenční abstrakty
Citace
Popis Isoelectric focusing (IEF) is an efficient method for separation of macromolecules. Classic IEF formats as capillary or gel are not suited for a processing of high volume of a sample. This disadvantage is solved by free flow IEF. Thanks to continuous flow of sample and ongoing separation the technique could be conveniently used for pre-separation and possibly separation of analytes. Method of divergent flow IEF (DF-IEF) invented in our department further improves separation efficiency of current free flow IEF instrument pattern. Based on our previous research we used the DF-IEF for concentration and purification of caseinomacropeptide (CMP). CMP originates during cheese making process by cleavage of kappa-casein with chymosin. It is heterogeneous group of differently glycated polypeptides based on the same primary amino acid structure. We used sweet bovine whey as a source of CMP since it composes about 25 % of whole whey protein. Particular focused fractions originated from sweet bovine whey were analyzed by capillary HPLC with C18 reverse phase column and UV detection. Presence of CMP in fractions was confirmed by comparison of obtained chromatograms with results of HPLC analysis of CMP standard. Results showed an ability of device to separate CMP from alpha-lactalbumin and betha-lactoglobulin. We proved that DF-IEF may be used for CMP purification as a technique alternative to ultrafiltration and ion-exchange chromatography.