Affinity immobilized metal resins
OwnerTran Quang Minhinventor
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Abstract
Pentadentate chelators (PDC) resins are the metal chelate resins capable of forming the octahedral complexes with several polyvalent metal ions including Cu2+, Ni2+, Zn2+ and Co2+ with five coordination sites occupied by the chelator. This results in the best stability of the complexes and in one coordination site free for interaction and selective binding of accessible cysteine/histidine residues and chiefly histidine containing biomolecules such as proteins or peptides etc. Cu-PDC can be used as concentration resins to reduce the volume of a protein solution. It can be used also as a universal support for immobilizing covalently all proteins, using a soluble carbodiimide.
Claims12
A pentadentate chelator resin having the formula:
- Resin-N (CH2—COOH)—(CH2)4—CH(COOH)—N(CH2—COOH)2, wherein said resin has pores which do not allow penetration by proteins having a molecular weight higher than 5000 Daltons.
A pentadentate chelator kit (PDC-kit) comprising four separated pentadentate chelator resins, wherein each of said resins is coordinated to a different divalent metal ion, wherein the divalent metal ions are Cu2+, Zn2+, Ni2+, and Co2+.
A method for the preparation of a column for covalently immobilizing a protein, said column comprising a pentadentate chelator (PDC) resin having the formula Resin-N (CH2—COOH)—(CH2)4—CH(COOH)—N(CH2—COOH)2, comprising:
- reacting an appropriately activated carrier matrix of a porous resin with a metal chelator having the formula:
- Bis-Lysine-Cu2+, in order to form a compound having the formula:
- Resin-ω-N-Lysine;
- reacting said compound having the formula:
- Resin-ω-N-Lysine, to form the pentadentate chelator resin;
- and
- contacting said resin with a composition comprising said protein.
A method for covalent immobilization of a protein comprising:
- contacting the protein with the pentadentate chelator resin having the formula Resin-N (CH2—COOH)—(CH2)4—CH(COOH)—N(CH2—COOH)2, wherein the pentadentate chelator resin is coordinated with divalent metal Cu2+, said contacting thereby forming a protein-PDC Cu2+ resin complex;
- contacting the protein-PDC Cu2+ resin complex with a water soluble carbodiimide;
- and
- removing the divalent metal Cu2+ from the protein-PDC Cu2+ resin complex.
A method for reducing the volume of a protein solution, comprising:
- contacting the protein with the pentadentate chelator resin having the formula Resin-N (CH2—COOH)—(CH2)4—CH(COOH)—N(CH2—COOH)2, wherein the pentadentate chelator resin is porous and coordinated with divalent metal Cu2+, said contacting thereby forming a protein-PDC Cu2+ resin complex;
- and
- eluting the protein from the PDC Cu2+ resin.
Dependencies are inferred from the claim text.