Method of preparing biologically active reagents from succinimide-containing polymers, analytical element and methods of use
AssigneeEastman Kodak Company
InventorsRichard C. Sutton, Ignazio S. Ponticello, Susan J. Danielson, Marsha D. B. Oenick
Life of this patent
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Abstract
Biologically active reagents are prepared from particles of copolymers having highly active succinimid groups. The reagents are prepared by covalently attaching biologically active substances, for example antibodies, to the particles, directly or indirectly through amide groups by displacement of highly active succinimid groups on the particle surface. These reagents are used to advantage in analytical elements, methods for the detection of specific binding ligands (such as immunological species) and immunoassays, and in purification methods such as affinity chromatography.
Claims11
A method of preparing a biologically active reagent comprising the step of reacting:
- (I) a water-insoluble, nonporous particle of a noncrosslinked copolymer having recurring units derived from:
- (a) from 0 to about 99.9 mole percent of one or more ethylenically unsaturated polymerizable oleophilic monomers which provide hydrophobicity to said copolymer, provided that none of said monomers are crosslinking monomers,
- (b) from about 0.1 to 100 mole percent of one or more ethylenically unsaturated polymerizable monomers having a succinimidoxycarbonyl group, and
- (c) from 0 to about 10 mole percent of one or more other ethylenically unsaturated polymerizable monomers, with
- (II) a biologically active substance to covalently attach said biologically active substance to said particle through an amide group by displacement of the succinimidoxy portion of said succinimidoxycarbonyl group.
An analytical element comprising a fluid-permeable substrate having one or more reaction zones therein, and containing in at least one of said zones, a biologically active reagent comprising:
- (I) a water-insoluble, nonporous particle of a noncrosslinked copolymer having recurring units derived from:
- (a) from 0 to about 99.9 mole percent of one or more ethylenically unsaturated polymerizable oleophilic monomers which provide hydrophobicity to said copolymer, provided that none of said monomers are crosslinking monomers,
- (b) from about 0.1 to 100 mole percent of one or more ethylenically unsaturated polymerizable monomers having a succinimidoxycarbonyl group, and
- (c) from 0 to about 10 mole percent of one or more other ethylenically unsaturated polymerizable monomers, and
- (II) a biologically active substance having been covalently attached to said particle through an amide group by displacement of the succinimidoxy portion of said succinimidoxycarbonyl group.
An analytical element comprising a nonporous support, having imposed thereon, in order and in fluid contact,
- a reagent layer containing one or more reagents for providing a detectable signal in response to an enzyme,
- a water-soluble layer containing an analog of a ligand labeled with said enzyme, and
- a porous spreading layer containing a biologically active reagent comprising:
- (I) a water-insoluble, nonporous particle of a noncrosslinked copolymer having recurring units derived from:
- (a) from 0 to about 99.9 mole percent of one or more ethylenically unsaturated polymerizable oleophilic monomers which provide hydrophobicity to said copolymer, provided that none of said monomers are crosslinking monomers,
- (b) from about 0.1 to 100 mole percent of one or more ethylenically unsaturated polymerizable monomers having a succinimidoxycarbonyl group, and
- (c) from 0 to about 10 mole percent of one or more other ethylenically unsaturated polymerizable monomers, and
- (II) a receptor specific for said ligand having been covalently attached to said particle through an amide group by displacement of the succinimidoxy portion of said succinimidoxycarbonyl group.
Dependencies are inferred from the claim text.