Comparison of anion-exchange and hydroxyapatite displacement chromatography for the isolation of whey proteins.
In displacement chromatography, several substances may be isolated and concomitantly concentrated, which makes this separation procedure attractive for the processing of diluted product streams containing a number of high value substances. Here, the suitability of anion-exchange and hydroxyapatite displacement chromatography for the processing of technical dairy whey is investigated. The pH and flow-rate of the carrier, the displacer chemistry and, in case of the apatite, the particle diameter of the stationary phase are considered. As a consequence of the pH sensitivity of the beta-lactoglobulin, one major whey component, apatite displacement chromatography is less then successful in whey separation. At a denaturing carrier pH (> 8.5) the beta-lactoglobulin zone is broad and stretches over the entire displacement train. At a lower carrier pH, previously successful polyanionic displacers do not bring about separation, while low-molecular-mass ones do, but they tend to overrun and thus contaminate the protein zones. In the case of anion-exchange displacement chromatography, polyanions, especially polyacrylic acid (PAA, M(r) 6000), constitute suitable displacers. Here too, a carrier pH of 8.0 is most suited to the separation of the whey proteins. The low-molecular mass-displacer iminodiacetic acid (IDA, M(r) 133.4), on the other hand, displaces only alpha-lactalbumin. The beta-lactoglobulin remains on the column. PAA is used as the displacer to process a dairy whey sample.
Vogt
S
S
Laboratoire de Biotechnologie Cellulaire et Moléculaire, Institute de Génie Chimique, ETH Lausanne, Switzerland.
Freitag
R
R
D000818:Animals; D000837:Anion Exchange Resins / Q000737:chemistry*; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D006882:Hydroxyapatites / Q000737:chemistry*; D007768:Lactalbumin / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D007782:Lactoglobulins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D008863:Microspheres; D008894:Milk Proteins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D010316:Particle Size; D011506:Proteins / Q000032:analysis; D000067816:Whey Proteins
D000837:Anion Exchange Resins; D006882:Hydroxyapatites; D007782:Lactoglobulins; D008894:Milk Proteins; D011506:Proteins; D000067816:Whey Proteins; D007768:Lactalbumin