Chelator and use thereof
The present invention relates to dimeric pentadentate chelators with exceptionally strong binding of metal ions, for detection, immobilization and purification of biomolecules. Dimeric chelators offer a cooperativity of binding of two adjacent immobilized metal ions simultaneously to a histidine-tagged biomolecule, which gives advantageous properties regarding strength of binding compared to a corresponding monomer chelator. In addition, a dimer increases the selectivity (ease of separation) against non-tagged biomolecules with low metal-ion affinity.
1. A dimeric pentadentate chelator having the following general formula:
wherein Q is a solid phase, a detection label, or a biomarker,
wherein Sc is a scaffold that contains at least two functional groups that couple two pentadentate chelators (PD), wherein Sc comprises diethylenetriamine, and
wherein PD is a pentadentate chelator having the formula:
2. The dimeric pentadentate chelator of claim 1 , wherein Q is a solid phase comprising a natural or synthetic polymer.
3. The dimeric pentadentate chelator of claim 2 , wherein the solid phase is a porous chromatographic support.
4. The dimeric pentadentate chelator of claim 2 , wherein Q is made of a cross-linked carbohydrate material.
5. The dimeric pentadentate chelator of claim 4 , wherein the cross-linked carbohydrate material is agarose, agar, cellulose, dextran, chitosan, konjac, carrageenan, gellan, or alginate.
6. The dimeric pentadentate chelator of claim 4 , wherein the cross-linked carbohydrate material is agarose.
7. The dimeric pentadentate chelator of claim 2 , wherein Q is made of one or more synthetic polymers.
8. The dimeric pentadentate chelator of claim 7 , wherein the one or more synthetic polymers comprise styrene derivatives, divinylbenzene, acryl amides, acrylate esters, methacrylate esters, vinyl esters, vinyl amides, or a combination thereof.
9. The dimeric pentadentate chelator of claim 2 , wherein Q comprises magnetic particles.
10. The dimeric pentadentate chelator of claim 1 , wherein Q is a solid phase comprising a sensor surface.
11. A dimeric pentadentate chelator having the following general formula:
wherein Q is a solid phase, a detection label, or a biomarker,
wherein Sc is a scaffold that contains at least two functional groups that couple two pentadentate chelators (PD), wherein Sc comprises diethylenetriamine,
wherein PD is a pentadentate chelator having the formula:
and
wherein Sp is a spacer provided between the scaffold Sc and the group Q, wherein the spacer has the following formula Xf-SP-Yf before coupling to Q and Sc, wherein Xf and Yf are selected from —OH, —SH or —OCH 2 CH═CH 2 , —CO 2 H, —NH 2 or NH—, —N 3 , CN, OTs, OMs, Cl, Br, —CHO, —C≡CH or epoxide groups, and wherein SP is a straight or branched configuration of 2-100 atoms comprising C, N, O, S and H.
12. The dimeric pentadentate chelator of claim 11 , wherein Xf is —CO 2 H and Yf is —SH.
13. The dimeric pentadentate chelator of claim 11 , wherein the spacer is derived from mercapto hexanoic acid.