Biologically active composition and the use thereof

• US 4,061,466 A
• Filed: 10/14/1975
• Issued: 12/06/1977
• Est. Priority Date: 10/16/1974
• Status: Expired due to Term

• Current Assignee: Bo Magnus Ekman, Ingvar Gosta Holger Sjoholm, Nils Roger Lindmark
• Original Assignee: Bo Magnus Ekman, Ingvar Gosta Holger Sjoholm, Nils Roger Lindmark
• Inventors: Sjoholm, Ingvar Gosta Holger, Lindmark, Nils Roger, Ekman, Bo Magnus
• Primary Examiner(s): Marantz, Sidney
• Application Number:
  US05/621,888
• Time in Patent Office: 784 Days
• Field of Search: 23/230 B, 23/230.3, 23/230.6, 195/63, 195/68, 195/DIG. 11, 424/1, 424/1.5, 424/12, 260/112 R, 252/184, 252/316
• US Class Current: 436/535
• CPC Class Codes: A61K 9/1635 obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates
• CPC Class Codes: A61K 9/167 with an outer layer or coating comprising drug; with chemically bound drugs or non-active substances on their surface with further drug-free outer coating A61K9/5073
• CPC Class Codes: B01J 13/14 Polymerisation; cross-linking
• CPC Class Codes: C12Q 1/002 Electrode membranes
• CPC Class Codes: G01N 33/549 with antigen or antibody entrapped within the carrier
• CPC Class Codes: Y10S 436/816 Alkaloids, amphetamines, and barbiturates
• CPC Class Codes: Y10S 436/828 Protein A
• CPC Class Codes: Y10S 530/813 Carrier is a saccharide
• CPC Class Codes: Y10S 530/817 Entrapped within the carrier, e.g. gel, hollow fibre

Abstract

Immobilized biologically active substances comprising microporous, spherical particles in gel form and in the form of a three-dimensional network. These particles contain the biologically active substance entrapped in the meshes of the network. The particles have sizes below 10 μm and the average diameter of the particles is in the range of from 0.5 to 4 μm, preferably 1 μm. The biologically active substance is capable of exerting its biological activity against substances which cannot penetrate into the meshes of the network of the particles. Preferably, the particles are gel grains obtained by copolymerization of acrylic compounds with cross-linkers. The immobilized biologically active substances are used for labelling or separation of cells, cell fragments, viruses or tissue structures having specific receptors or of substances having specific surface structures.

Claims

1. A biologically active composition comprising microporous, spherical particles in gel form, said particles having a three-dimensional polymeric network, diameters less than 10 μ

m and an average diameter of from 0.5 to 4μ

m and a biologically active macromolecular substance immobilized within the meshes of said polymeric network but retainining its biological activity.

2. A biologically active composition according to claim 1 wherein the spherical particles consist of a copolymer of a low-molecular acrylic compound or of 1-vinyl-2-pyrrolidinone with a low-molecular divinyl compound.

3. A biologically active composition according to claim 2 wherein the spherical particles consist of a copolymer of an acrylamide of the formula ##STR2## wherein R₁ is hydrogen or methyl and R₂ is OH or --NHR₃, wherein R₃ is hydrogen or a straight or branched alkyl group of 1-6 carbon atoms, which alkyl group is unsubstituted or substituted with a hydroxyl or an oxo group, with N,N'"'"'-methylene-bis-acrylamide.

4. A process for carrying out a bioassay for quantitatively determining a particular substance in a biological sample comprising adding to said sample a reagent comprising of a biologically active composition comprising

a. microporous, spherical particles in gel form, said particles having a three-dimensional polymeric network, diameters less than 10 μ

m and an average diameter of from 0.5 to 4 μ

m and

b. a biologically active macromolecular substance immobilized within the meshes of said polymeric network but retaining its biological activity, said biologically active macromolecular substance being specific to said particular substance.

5. Process according to claim 4 wherein the average particle diameter is 1 μ

m.

6. Process according to claim 4 wherein the biologically active macromolecular substance is labelled with an atom or group capable of emitting radiation or with an enzyme or a group producing a color.

7. A process for labelling a biological member selected from the group consisting of cells, cell fragments, viruses, tissue structures having specific receptors and substances having specific surface conformations comprising contacting said member with a reagent comprising a biologically active composition comprising

a. microporous, spherical particles in gel form, said particles having three-dimensional polymeric network, diameters less than 10 μ

m and an average diameter of from 0.5 to 4 μ

m and

b. a biologically active macromolecular substance immobilized within the meshes of said polymeric network but retainings its biological activity, said biologically active macromolecular substance being specific to said member and being labelled with a radioactive atom, an enzyme or fluorophore.

8. Process according to claim 7 wherein the average particle diameter is 1 μ

m.

9. A method of separating a biological member selected from the group consisting of cells, cell fragments, viruses, tissue structures having specific receptors and substances having specific surface conformtions from a biological sample containing said member comprising mixing said biological sample with a reagent comprising a biologically active composition comprising

11. Method according to claim 9, wherein the average particle diameter is 1 μ

m.

12. Method according to claim 9 wherein the biologically active macromolecular substance is labelled with an atom or group capable of emitting radiation or with an enzyme or a group producing a color.

10. a. microporous, spherical particles in gel form, said particles having a three-dimensional polymeric network, diameters less than 10 μ

m and an average diameter of from 0.5 to 4 μ

m and

b. a biologically active macromoleeular substance immobilized within the meshes of said polymeric network but retaining its biological activity, said biologically active macromolecular substance being specific to said member and separating the resulting conjugate of said member-said reagent by physical means.