US4829098A
Expired / Lapsed
US
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Immobilized biomolecules and method of making same
Assignee: University of Washington
•
Filing: 1986-06-19
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Granted: 1989-05-09
•
Expires: 2006-06-19
Abstract
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 DEG C. in air, while the grafting step is carried out at 0 DEG 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.
Inventors
- 👤 Allan S. Hoffman
- 👤 Liang C. Dong
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 14:01
CPC Classifications
C; C12; C12N; C12N11/00; C12N11/02; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54393; G01N33/544; G01N33/545; G01N2650/00
Claims Analysis
Total Claims: 54
Claim 1
Independent Claim
71 words
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° C. to about -78° C.; and contacting the irradiated substrate with a biomolecule and free monomer in aqueous solution at about 0° C. in a substantially oxygen-free atmosphere to form a graft copolymer wherein the biomolecule is immobilized at least in part within the graft copolymer.
Claim 2
Independent Claim
71 words
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° C. to about -78° C.; and contacting the irradiated substrate with a biomolecule and free monomer in aqueous solution at about 0° C. in a substantially oxygen-free atmosphere to form a graft copolymer wherein the biomolecule is immobilized at least in part within the graft copolymer.
Claim 3
Dependent Claim
11 words
The method of claim 1 wherein the biomolecule is an enzyme.
Claim 4
Dependent Claim
10 words
The method of claim 2 wherein the enzyme is asparaginase.
Claim 5
Dependent Claim
11 words
The method of claim 1 wherein the biomolecule is a catalyst.
Claim 6
Dependent Claim
11 words
The method of claim 1 wherein the biomolecule is an antibody.
Claim 7
Dependent Claim
11 words
The method of claim 1 wherein the biomolecule is an antigen.
Claim 8
Dependent Claim
11 words
The method of claim 1 wherein the biomolecule is a drug.
Claim 9
Dependent Claim
11 words
The method of claim 1 wherein the biomolecule is a hormone.
Claim 10
Dependent Claim
11 words
The method of claim 1 wherein the biomolecule is a pesticide.
Claim 11
Dependent Claim
11 words
The method of claim 1 wherein the biomolecule is a fertilizer.
Claim 12
Dependent Claim
11 words
The method of claim 11 wherein the biomolecule is an insecticide.
Claim 13
Dependent Claim
11 words
The method of claim 1 wherein the biomolecule is a dye.
Claim 14
Dependent Claim
11 words
The method of claim 1 wherein the biomolecule is a vitamin.
Claim 15
Dependent Claim
19 words
The method of claim 1 wherein the biomolecule is a single or double stranded sequence of DNA or RNA.
Claim 16
Dependent Claim
12 words
The method of claim 1 wherein the biomolecule is a nucleic acid.
Claim 17
Dependent Claim
10 words
The method of claim 1 wherein the substrate is hydrophilic.
Claim 18
Dependent Claim
11 words
The method of claim 1 wherein the substrate is a polysaccharide.
Claim 19
Dependent Claim
21 words
The method of claim 1 wherein the substrate is selected from the group consisting of cellulose, agarose, sepharose and cross-linked dextran.
Claim 20
Dependent Claim
14 words
The method of claim 1 wherein the source of ionizing radiation is 60 Co.
Claim 21
Dependent Claim
13 words
The method of claim 1 wherein the source of ionizing radiation is X-rays.
Claim 22
Dependent Claim
14 words
The method of claim 1 wherein the source of ionizing radiation is high-energy electrons.
Claim 23
Dependent Claim
17 words
The method of claim 1 wherein the source of ionizing radiation is radio frequency plasma gas discharge.
Claim 24
Dependent Claim
16 words
The method of claim 1 wherein a substrate is exposed to ionizing radiation at -78° C.
Claim 25
Dependent Claim
17 words
The method of claim 1 wherein the monomer is characterized by having at least one vinyl group.
Claim 26
Dependent Claim
17 words
The method of claim 1 wherein the monomer is characterized by having at least one allyl group.
Claim 27
Dependent Claim
17 words
The method of claim 1 wherein the monomer is selected from the group shown in Table 1.
Claim 28
Independent Claim
73 words
A method of producing an immobilized biomolecule comprising the steps of: exposing a hydrophilic solid-phase polymeric substrate to ionizing radiation at a temperature between about 0° C. to about -78° C.; and contacting the irradiated substrate with a monomer-conjugated biomolecule and free monomer in aqueous solution at about 0° C. in a substantially oxygen-free atmosphere to form a graft copolymer, wherein the biomolecule is immobilized at least in part within the graft copolymer.
Claim 29
Independent Claim
73 words
A method of producing an immobilized biomolecule comprising the steps of: exposing a hydrophilic solid-phase polymeric substrate to ionizing radiation at a temperature between about 0° C. to about -78° C.; and contacting the irradiated substrate with a monomer-conjugated biomolecule and free monomer in aqueous solution at about 0° C. in a substantially oxygen-free atmosphere to form a graft copolymer, wherein the biomolecule is immobilized at least in part within the graft copolymer.
Claim 30
Dependent Claim
11 words
The method of claim 27 wherein the biomolecule is an enzyme.
Claim 31
Dependent Claim
10 words
The method of claim 28 wherein the enzyme is asparaginase.
Claim 32
Dependent Claim
11 words
The method of claim 27 wherein the biomolecule is a catalyst.
Claim 33
Dependent Claim
11 words
The method of claim 27 wherein the biomolecule is an antibody.
Claim 34
Dependent Claim
11 words
The method of claim 27 wherein the biomolecule is an antigen.
Claim 35
Dependent Claim
11 words
The method of claim 27 wherein the biomolecule is a drug.
Claim 36
Dependent Claim
11 words
The method of claim 27 wherein the biomolecule is a hormone.
Claim 37
Dependent Claim
11 words
The method of claim 27 wherein the biomolecule is a pesticide.
Claim 38
Dependent Claim
11 words
The method of claim 27 wherein the biomolecule is a fertilizer.
Claim 39
Dependent Claim
11 words
The method of claim 27 wherein the biomolecule is an insecticide.
Claim 40
Dependent Claim
11 words
The method of claim 27 wherein the biomolecule is a dye.
Claim 41
Dependent Claim
11 words
The method of claim 27 wherein the biomolecule is a vitamin.
Claim 42
Dependent Claim
19 words
The method of claim 27 wherein the biomolecule is a single or double stranded sequence of DNA or RNA.
Claim 43
Dependent Claim
12 words
The method of claim 27 wherein the biomolecule is a nucleic acid.
Claim 44
Dependent Claim
10 words
The method of claim 27 wherein the substrate is hydrophilic.
Claim 45
Dependent Claim
11 words
The method of claim 27 wherein the substrate is a polysaccharide.
Claim 46
Dependent Claim
21 words
The method of claim 27 wherein the substrate is selected from the group consisting of cellulose, agarose, sepharose and cross-linked dextran.
Claim 47
Dependent Claim
14 words
The method of claim 27 wherein the source of ionizing radiation is 60 Co.
Claim 48
Dependent Claim
13 words
The method of claim 27 wherein the source of ionizing radiation is X-rays.
Claim 49
Dependent Claim
14 words
The method of claim 27 wherein the source of ionizing radiation is high-energy electrons.
Claim 50
Dependent Claim
18 words
The method of claim 27 wherein the source of ionizing radiation is a radio frequency plasma gas discharge.
Claim 51
Dependent Claim
16 words
The method of claim 27 wherein a substrate is exposed to ionizing radiation at -78° C.
Claim 52
Dependent Claim
17 words
The method of claim 27 wherein the monomer is characterized by having at least one vinyl group.
Claim 53
Dependent Claim
17 words
The method of claim 27 wherein the monomer is characterized by having at least one allyl group.
Claim 54
Dependent Claim
17 words
The method of claim 27 wherein the monomer is selected from the group shown in Table 1.
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