Protein separation with magnetic adsorbents in micellar aqueous two-phase systems
Becker, J. / Thomas, O. / Franzreb, M. (2008)
Separation and Purification Technology 65 (2008), 1, 46–53
- Datum: 2008
Becker, J. / Thomas, O. / Franzreb, M. (2008). „Protein separation with magnetic adsorbents in micellar aqueous two-phase systems“. In: Separation and Purification Technology 65 (2008), 1, 46–53
We introduce a new concept for the purification and concentration of macromolecules such as proteins based on the use of magneticadsorbents in concert with micellaraqueoustwo-phasesystems. Aqueous solutions of certain nonionic surfactants can, depending on the prevailing temperature, separate into two co-existing phases, one enriched, the other depleted in surfactant.
These are known as micellaraqueoustwo-phasesystems. Hydrophilic proteins are normally excluded from the ‘surfactant-rich’ phase of micellaraqueoustwo-phasesystems. We show here that by introducing magnetic microadsorbents into micellaraqueoustwo-phases the phase selectivity of a hydrophilic protein can be effectively inverted.
The magneticadsorbents adsorb the target protein and pull it out of the ‘surfactant-depleted’ phase into the ‘surfactant-rich’ phase from which it is normally excluded. Starting from a model solution containing a 1:1 ratio of the hen egg white proteins lysozyme and ovalbumin, the application of magnetic cation exchange micro-adsorbents combined with the nonionic surfactant Triton X-114 resulted in a target lysozyme yield of 74% and purity of >80%.
The combination of selective magneticadsorbents with micellaraqueoustwo-phasesystems yields a highly tunable extraction system. Potentially, these so-called ‘Magnetic Extraction Phases’ provide the basis for flexible and easily scaled fractionation processes for hydrophilic proteins.