Immobilized biomolecules and method of making same
AssigneeThe Board of Regents of the University of Washington
InventorsAllan S. Hoffman, Liang C. Dong
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Abstract
A method of biomolecule immobilization is described in which the biomolecule itself or, alternatively, a monomer-conjugated biomolecule, is grafted with free monomer onto a hydrophilic, solid-phase, polymeric substrate which has been pre-irradiated with ionizing radiation. The pre-irradiation step is carried out, preferably at -78.degree. C. in air, while the grafting step is carried out at 0.degree. C. in a substantially oxygen-free atmosphere. The technique is applicable to immobilization of a wide variety of biomolecules, such as enzymes, catalysts, hormones, lectins, drugs, vitamins, antibodies, antigens, nucleic acids, DNA and RNA segments, pesticides, dyes and fertilizers. The products may be used for therapeutic or diagnostic applications or bioseparations.
Claims52
A method of producing an immobilized biomolecule comprising the steps of:
- exposing a solid-phase polymeric substrate to ionizing radiation at a temperature between about 0.degree. C. to about -78.degree. C.;
- and
- contacting the irradiated substrate with a biomolecule and free monomer, in aqueous solution in a substantially oxygen-free atmosphere at a temperature less than the temperature at which the biomolecule inactivates, to form a graft copolymer, wherein the biomolecule is immobilized at least in part within the graft copolymer.
A method of producing an immobilized biomolecule comprising the steps of:
- exposing a solid-phase polymeric substrate to ionizing radiation at a temperature between about 0.degree. C. to about -78.degree. C.;
- and
- contacting the irradiated substrate with a monomer-conjugated biomolecule and free monomer, in aqueous solution in a substantially oxygen-free atmosphere at a temperature less than the temperature at which the biomolecule inactivates, to form a graft copolymer, wherein the biomolecule is immobilized at least in part within the graft copolymer.
The method of claim 278 wherein the monomer is selected from the group shown in Table 1.
Dependencies are inferred from the claim text.