Use of ph-responsive polymers
The present invention relates to a method of isolating target compounds from a liquid, which comprises at a first pH, contacting the liquid with a separation medium that exhibits surface-localised pH-responsive polymers in to adsorb the target compound via hydrophobic interactions; and adding an eluent, which is of a second pH and provides a conformational change of said pH-responsive polymers to release said compounds. The elution is advantageously performed by a pH gradient and/or by a salt gradient.
1 . A method of isolating at least one target compound from a liquid, which method comprises the steps of
(a) contacting the liquid, at a first pH, with a separation medium that includes surface-localised pH-responsive polymers to adsorb the target compound(s) via hydrophobic interactions; and
(b) adding an eluent of a second pH value, which eluent provides a conformational change of said pH-responsive polymers, to release the target compound(s) from the separation medium.
2 . The method of claim 1 , wherein the second pH value is lower than the first pH value.
3 . The method of claim 2 , wherein the eluent comprises a decreasing pH gradient.
4 . The method of claim 1 , wherein the conductivity of the eluent differs from the conductivity of the liquid of step (a), while the second pH value is essentially equal to the first pH value.
5 . The method of claim 1 , wherein the eluent comprises a salt gradient.
6 . The method of claim 1 , wherein in step (a), the separation medium is uncharged.
7 . The method of claim 1 , wherein in step (a), the target compound(s) are adsorbed also by additional interactions between pH-responsive polymers and target compounds, said additional interactions being selected from the group consisting of charge-charge interactions, van der Waals interactions and interactions based on cosolvation/cohydration.
8 . The method of claim 1 , wherein in step (b), the separation medium is uncharged.
9 . The method of claim 1 , wherein in step (b), the pH-responsive polymers are less hydrophobic than in step (a).
10 . The method of claim 1 , wherein the conformational change of the polymers is provided by polymer self-association and/or polymer association with the medium.
11 . The method of claim 1 , wherein the pH-responsive polymers are copolymers.
12 . The method of claim 1 , wherein each pH-responsive polymer includes a hydrophobic part, a hydrophilic part and a pH-responsive part.
13 . The method of claim 1 , wherein the pH-responsive polymers include pendant pH-sensitive groups selected from the group consisting of —COOH groups; —OPO(OH) 2 groups; —SO 3 − groups; SO 2 NH 2 groups; —CNH 2 groups —C 2 NH groups; and —C 3 N groups.
14 . The method of claim 1 , wherein the target compound is a biomolecule.
15 - 18 . (canceled)
19 . A hydrophobic interaction chromatography (HIC) medium, which is comprised of a matrix to which surface-localised pH-responsive polymers have been attached, which polymers include HIC ligands.
20 . The medium of claim 19 , wherein the pH-responsive groups of the polymers have been selected from the group consisting of —COOH groups; —OPO(OH) 2 groups; —SO 3 — groups; SO 2 NH 2 groups; —CNH 2 groups —C 2 NH groups; and —C 3 N groups.
21 . A kit for isolating target compounds, which kit comprises, in separate compartments, a chromatography column packed with a medium comprised of a matrix to which surface-localised pH-responsive polymers, which exhibit HIC ligands, have been attached; an adsorption buffer of a first pH; and an eluent of a second pH, which is lower that said first pH.
22 . The kit of claim 21 , wherein the pH-responsive groups of the polymers have been selected from the group consisting of —COOH groups; —OPO(OH) 2 groups; —SO 3 — groups; SO 2 NH 2 groups; —CNH 2 groups —C 2 NH groups; and —C 3 N groups.