Magnetic polymer particles
A conjugate comprising a magnetic polymer particle bound to a carboxymethylated aspartate chelating ligand, optionally chelating a metal ion.
1 . A conjugate comprising a magnetic polymer particle bound to a carboxymethylated aspartate chelating ligand.
2 . A conjugate comprising a magnetic polymer particle bound to a carboxymethylated aspartate ligand chelating a metal atom or ion.
3 . A conjugate as claimed in claim 2 wherein said metal is a transition metal or a metal of group 13.
4 . A conjugate as claimed in claim 3 wherein said metal is Ni, Fe, Ga, Mn, Co, Cu and Zn.
5 . A conjugate as claimed in claim 4 wherein said metal is Fe, Ga, Mn and Co.
6 . A conjugate as claimed in claim 2 wherein said metal is in the 2+ or 3+ oxidation state.
7 . A conjugate as claimed in claim 6 wherein said metal is Co 2+ , Fe 3+ , Ga 3+ and Cu 2+ .
8 . A conjugate as claimed in claim 7 wherein said metal is Co 2+ .
9 . A conjugate as claimed in claim 1 wherein there are at least three atoms between the nitrogen atom of the carboxymethylated aspartate ligand and the particle surface.
10 . A conjugate as claimed in claim 9 being of formula
(MPP=magnetic polymer particle)
wherein the wavy line represents a 3 to 20 atom linker selected from NH-alkylene, NH—CO-alkylene, O-alkylene, OCO-alkylene, S-alkylene or SCO-alkylene.
11 . A conjugate as claimed in claim 10 wherein the wavy line represents NH—C 5 H 12 — or NH—C 6 H 13 —.
12 . A conjugate as claimed in claim 1 wherein said polymer comprises a cross-linked styrene divinyl benzene polymer.
13 . A conjugate as claimed in claim 1 wherein the magnetic polymer particle has a diameter of 0.5 to 8 μm.
14 . A conjugate as claimed in claim 12 wherein said magnetic polymer particle has a diameter of 0.8 to 1.2 μm.
15 . A conjugate as claimed in claim 1 being uncharged.
16 . A conjugate as claimed in claim 2 additionally chelated to a histidine tagged recombinant protein/peptide, His, Cys, Met, GIn, Asn, Lys and/or Tyr residue containing native protein/peptide or phosphorylated protein/peptide.
17 . A conjugate as claimed in claim 2 additionally chelated to a histidine tagged recombinant protein/peptide.
18 . A conjugate as claimed in claim 16 characterised in that where said conjugate binds a phosphorylated protein/peptide, said metal is Fe or Ga.
19 . A process for the preparation of a conjugate comprising a magnetic polymer particle bound to a Cm-Asp ligand comprising reacting a Cm-Asp ligand of formula (II)
(wherein each R independently represents hydrogen or a protecting group and X represents a 2 to 20 atom group) with a magnetic polymer particle, and optionally coordinating the resulting conjugate to a metal atom or ion.
20 . A compound of formula (II)
(wherein each R independently represents hydrogen or a protecting group and X represents a 2 to 20 atom group) or an analogue therefore in which the R groups are absent and a metal chelated.
21 . A compound as claimed in claim 20 wherein X is a C5 or C6-alkylene group.
22 . A compound of formula (III) or an analogue thereof in which a metal is
23 . A process for the preparation of a compound of formula
(wherein each R independently represents hydrogen or a protecting group and X represents an C 2-20 alkylene linker);
comprising reacting a compound of formula Hal-X 1 —CN (wherein Hal is a halide and X 1 represents an C 1-19 alkylene linker) with a compound of formula
(wherein Pr is a protecting group)
reacting the resulting product with a compound of formula Hal-CH 2 COOPr to form a compound
reducing the nitrile to an amino group; and optionally deprotecting the carboxyl groups.
24 . Use of a conjugate as claimed in claim 2 to 18 in an assay.
25 . Use of a conjugate as claimed in claim 2 to 18 in the purification of histidine tagged recombinant proteins/peptides, His, Cys, Met, Gin, Asn, Lys and/or Tyr residue containing native proteins/peptides or phosphorylated proteins/peptides.