US4481137A — Glycoproteins and processes for their production

Claim 1

Independent Claim 222 words

A substantially purified form of a glycoprotein (CB X ) produced from non-established cells or cells of established cell lines of reticulo-endothelial cells, lymphoblasts, leukemia cells or fibroblasts originated from warm-blooded animals, having an anti-tumor effect and having the following properties: (a) molecular weight: 12,000-17,000; (b) color reactions: it exhibits a color indicating proteins in the Lowry reaction, exhibits a color indicating peptide bonds and amino acids in the ninhydrin reaction after hydrolysis with hydrochloric acid, and exhibits a color indicating sugars in the phenol-sulfuric acid reaction, the anthrone-sulfuric acid reaction, the indole-sulfuric acid reaction and the tryptophane-sulfuric acid reaction; (c) appearance and solubility: white powder soluble in water, aqueous sodium chloride and phosphate buffer, and sparingly soluble in benzene, hexane and chloroform; (d) sugar content: sugar content is 27-33%, 17-20% of the total sugar being hexoses, 5-7% being hexosamines and 5-6% being sialic acids; (e) isoelectric point: 4.2-7.3; (f) adsorbability: adsorbable on Ulex europeus agglutinin-conjugated Sephadex in 0.01M phosphate buffer (pH 7.2); (g) stability: stable in an aqueous solution of pH 2.0, pH 7.0 or pH 11.0 at 4° C. for 24 hours or longer and in an aqueous solution of pH 7.0 at 60° C. for 3 hours or longer; (h) cytotoxicity: it selectively damages tumor cells without substantially damaging normal cells; and (i) differentiation: induces differentiation of tumor cells.

Claim 2

Dependent Claim 222 words

A substantially purified form of a glycoprotein (CB X ) produced from non-established cells or cells of established cell lines of reticulo-endothelial cells, lymphoblasts, leukemia cells or fibroblasts originated from warm-blooded animals, having an anti-tumor effect and having the following properties: (a) molecular weight: 12,000-17,000; (b) color reactions: it exhibits a color indicating proteins in the Lowry reaction, exhibits a color indicating peptide bonds and amino acids in the ninhydrin reaction after hydrolysis with hydrochloric acid, and exhibits a color indicating sugars in the phenol-sulfuric acid reaction, the anthrone-sulfuric acid reaction, the indole-sulfuric acid reaction and the tryptophane-sulfuric acid reaction; (c) appearance and solubility: white powder soluble in water, aqueous sodium chloride and phosphate buffer, and sparingly soluble in benzene, hexane and chloroform; (d) sugar content: sugar content is 27-33%, 17-20% of the total sugar being hexoses, 5-7% being hexosamines and 5-6% being sialic acids; (e) isoelectric point: 4.2-7.3; (f) adsorbability: adsorbable on Ulex europeus agglutinin-conjugated Sephadex in 0.01M phosphate buffer (pH 7.2); (g) stability: stable in an aqueous solution of pH 2.0, pH 7.0 or pH 11.0 at 4° C. for 24 hours or longer and in an aqueous solution of pH 7.0 at 60° C. for 3 hours or longer; (h) cytotoxicity: it selectively damages tumor cells without substantially damaging normal cells; and (i) differentiation: induces differentiation of tumor cells.

Claim 3

Dependent Claim 214 words

A substantially purified form of a glycoprotein (CB X1 ), produced from non-established cells or cells of established cell lines of reticulo-endothelial cells, lymphoblasts, leukemia cells or fibroblasts originated from warm-blooded animals, having an anti-tumor effect and having the following properties: (a) molecular weight: 70,000-90,000; (b) color reactions: it exhibits a color indicating proteins in the Lowry reaction, exhibits a color indicating peptide bonds and amino acids in the ninhydrin reaction after hydrolysis with hydrochloric acid, and exhibits a color indicating sugars in the phenol-sulfuric acid reaction, the anthrone-sulfuric acid reaction, the indole-sulfuric acid reaction and the tryptophane-sulfuric acid reaction; (c) appearance and solubility: white powder soluble in water, aqueous sodium chloride and phosphate buffer, and sparingly soluble in benzene, hexane and chloroform; (d) sugar content: sugar content is 35-45%, 23-28% of the total sugar being hexoses, 8-11% being hexosamines and 4-6% being sialic acids; (e) isoelectric point: 4.3-6.2; (f) adsorbability: adsorbable on Ulex europeus agglutinin-conjugated Sephadex in 0.01M phosphate buffer (pH 7.2); (g) stability: stable in an aqueous solution of pH 2.0, pH 7.0 pH 11.0 at 4° C. for 24 hours or longer and in an aqueous solution of pH 7.0 at 60° C. for 3 hours or longer; and (h) cytotoxicity: it selectively damages tumor cells without substantially damaging normal cells.

Claim 4

Dependent Claim 214 words

A substantially purified form of a glycoprotein (CB X1 ), produced from non-established cells or cells of established cell lines of reticulo-endothelial cells, lymphoblasts, leukemia cells or fibroblasts originated from warm-blooded animals, having an anti-tumor effect and having the following properties: (a) molecular weight: 70,000-90,000; (b) color reactions: it exhibits a color indicating proteins in the Lowry reaction, exhibits a color indicating peptide bonds and amino acids in the ninhydrin reaction after hydrolysis with hydrochloric acid, and exhibits a color indicating sugars in the phenol-sulfuric acid reaction, the anthrone-sulfuric acid reaction, the indole-sulfuric acid reaction and the tryptophane-sulfuric acid reaction; (c) appearance and solubility: white powder soluble in water, aqueous sodium chloride and phosphate buffer, and sparingly soluble in benzene, hexane and chloroform; (d) sugar content: sugar content is 35-45%, 23-28% of the total sugar being hexoses, 8-11% being hexosamines and 4-6% being sialic acids; (e) isoelectric point: 4.3-6.2; (f) adsorbability: adsorbable on Ulex europeus agglutinin-conjugated Sephadex in 0.01M phosphate buffer (pH 7.2); (g) stability: stable in an aqueous solution of pH 2.0, pH 7.0 pH 11.0 at 4° C. for 24 hours or longer and in an aqueous solution of pH 7.0 at 60° C. for 3 hours or longer; and (h) cytotoxicity: it selectively damages tumor cells without substantially damaging normal cells.

Claim 5

Dependent Claim 215 words

A substantially purified form of a glycoprotein (CB X2 ), produced from non-established cells or cells of established cell lines of reticulo-endothelial cells, lymphoblasts, leukemia cells or fibroblasts originated from warm-blooded animals, having an anti-tumor effect and having the following properties: (a) molecular weight: 40,000-50,000; (b) color reactions: it exhibits a color indicating proteins in the Lowry reaction, exhibits a color indicating peptide bonds and amino acids in the ninhydrin reaction after hydrolysis with hydrochloric acid, and exhibits a color indicating sugars in the phenol-sulfuric acid reaction, the anthrone-sulfuric acid reaction, the indole-sulfuric acid reaction and the tryptophane-sulfuric acid reaction; (c) appearance and solubility: white powder soluble in water, aqueous sodium chloride and phosphate buffer, and sparingly soluble in benzene, hexane and chloroform; (d) sugar content: sugar content is 30-37%, 20-23% of the total sugar being hexoses, 6-8% being hexosamines and 4-6% being sialic acids; (e) isoelectric point: 4.2-7.3; (f) adsorbability: adsorbable on Ulex europeus agglutinin-conjugated Sephadex in 0.01M phosphate buffer (pH 7.2); (g) stability: stable in an aqueous solution of pH 2.0, pH 7.0 or pH 11.0 at 4° C. for 24 hours or longer and in an aqueous solution of pH 7.0 at 60° C. for 3 hours or longer; and (h) cytotoxicity: it selectively damaged tumor cells without substantially damaging normal cells.

Claim 6

Dependent Claim 215 words

A substantially purified form of a glycoprotein (CB X2 ), produced from non-established cells or cells of established cell lines of reticulo-endothelial cells, lymphoblasts, leukemia cells or fibroblasts originated from warm-blooded animals, having an anti-tumor effect and having the following properties: (a) molecular weight: 40,000-50,000; (b) color reactions: it exhibits a color indicating proteins in the Lowry reaction, exhibits a color indicating peptide bonds and amino acids in the ninhydrin reaction after hydrolysis with hydrochloric acid, and exhibits a color indicating sugars in the phenol-sulfuric acid reaction, the anthrone-sulfuric acid reaction, the indole-sulfuric acid reaction and the tryptophane-sulfuric acid reaction; (c) appearance and solubility: white powder soluble in water, aqueous sodium chloride and phosphate buffer, and sparingly soluble in benzene, hexane and chloroform; (d) sugar content: sugar content is 30-37%, 20-23% of the total sugar being hexoses, 6-8% being hexosamines and 4-6% being sialic acids; (e) isoelectric point: 4.2-7.3; (f) adsorbability: adsorbable on Ulex europeus agglutinin-conjugated Sephadex in 0.01M phosphate buffer (pH 7.2); (g) stability: stable in an aqueous solution of pH 2.0, pH 7.0 or pH 11.0 at 4° C. for 24 hours or longer and in an aqueous solution of pH 7.0 at 60° C. for 3 hours or longer; and (h) cytotoxicity: it selectively damaged tumor cells without substantially damaging normal cells.

Claim 7

Dependent Claim 230 words

A substantially purified form of a glycoprotein (CB X3 ), produced from non-established cells or cells of established cell lines of reticulo-endothelial cells, lymphoblasts, leukemia cells or fibroblasts originated from warm-blooded animals, having an anti-tumor effect and having the following properties: (a) molecular weight: 7,000-9,000; (b) color reactions: it exhibits a color indicating proteins in the Lowry reaction, exhibits a color indicating peptide bonds and amino acids in the ninhydrin reaction after hydrolysis with hydrochloric acid, and exhibits a color indicating sugars in the phenol-sulfuric acid reaction, the anthrone-sulfuric acid reaction, the indole-sulfuric acid reaction and the tryptophane-sulfuric acid reaction; (c) appearance and solubility: white powder soluble in water, aqueous sodium chloride and phosphate buffer, and sparingly soluble in benzene, hexane and chloroform; (d) sugar content: sugar content is 8-15%, 6-10% of the total sugar being hexoses, 1-2% being hexosamines and 1-3% being sialic acids; (e) adsorbability: adsorbable on carboxymethylcellulose in an ion exchange chromatography in 0.05M phosphate buffer (pH 6.4) using carboxymethylcellulose; (f) stability: stable in an aqueous solution of pH 2.0, pH 7.0 or pH 11.0 at 4° C. for 24 hours or longer and in an aqueous solution of pH 7.0 at 60° C. for 3 hours or longer; (g) cytotoxicity: it selectively damages tumor cells without substantially damaging normal cells; and (h) the amino acid sequence of the N terminal of its protein portion is Alanine-Alanine-.

Claim 8

Dependent Claim 230 words

A substantially purified form of a glycoprotein (CB X3 ), produced from non-established cells or cells of established cell lines of reticulo-endothelial cells, lymphoblasts, leukemia cells or fibroblasts originated from warm-blooded animals, having an anti-tumor effect and having the following properties: (a) molecular weight: 7,000-9,000; (b) color reactions: it exhibits a color indicating proteins in the Lowry reaction, exhibits a color indicating peptide bonds and amino acids in the ninhydrin reaction after hydrolysis with hydrochloric acid, and exhibits a color indicating sugars in the phenol-sulfuric acid reaction, the anthrone-sulfuric acid reaction, the indole-sulfuric acid reaction and the tryptophane-sulfuric acid reaction; (c) appearance and solubility: white powder soluble in water, aqueous sodium chloride and phosphate buffer, and sparingly soluble in benzene, hexane and chloroform; (d) sugar content: sugar content is 8-15%, 6-10% of the total sugar being hexoses, 1-2% being hexosamines and 1-3% being sialic acids; (e) adsorbability: adsorbable on carboxymethylcellulose in an ion exchange chromatography in 0.05M phosphate buffer (pH 6.4) using carboxymethylcellulose; (f) stability: stable in an aqueous solution of pH 2.0, pH 7.0 or pH 11.0 at 4° C. for 24 hours or longer and in an aqueous solution of pH 7.0 at 60° C. for 3 hours or longer; (g) cytotoxicity: it selectively damages tumor cells without substantially damaging normal cells; and (h) the amino acid sequence of the N terminal of its protein portion is Alanine-Alanine-.

Claim 9

Dependent Claim 238 words

A process for producing a glycoprotein (CB X ) having an anti-tumor effect, comprising: culturing non-established cells or cells of established cell lines of reticulo-endothelial cells, lymphoblasts, leukemia cells or fibroblasts originated from warm-blooded animals, and isolating glycoproteins from a homogeneous mass or a cultured supernatant of said cultured cells, said isolated glycoproteins having the following properties: (a) molecular weight: 12,000-17,000; (b) color reactions: it exhibits a color indicating proteins in the Lowry reaction, exhibits a color indicating peptide bonds and amino acids in the ninhydrin reaction after hydrolysis with hydrochloric acid, and exhibits a color indicating sugars in the phenol-sulfuric acid reaction, the anthrone-sulfuric acid reaction, the indole-sulfuric acid reaction and the tryptophane-sulfuric acid reaction; (c) appearance and solubility: white powder soluble in water, aqueous sodium chloride and phosphate buffer, and sparingly soluble in benzene, hexane and chloroform; (d) sugar content: sugar content is 27-33%, 17-20% of the total sugar being hexoses, 5-7% being hexosamines and 5-6% being sialic acids; (e) isoelectric point: 4.2-7.3; (f) adsorbability: adsorbable on Ulex europeus agglutinin-conjugated Sephadex in 0.01M phosphate buffer (pH 7.2); (g) stability: stable in an aqueous solution of pH 2.0, pH 7.0 or pH 11.0 at 4° C. for 24 hours or longer and in an aqueous solution of pH 7.0 at 60° C. for 3 hours or longer; (h) cytotoxicity: it selectively damages tumor cells without substantially damaging normal cells; and (i) differentiation: induces differentiation of tumor cells.

Claim 10

Dependent Claim 238 words

A process for producing a glycoprotein (CB X ) having an anti-tumor effect, comprising: culturing non-established cells or cells of established cell lines of reticulo-endothelial cells, lymphoblasts, leukemia cells or fibroblasts originated from warm-blooded animals, and isolating glycoproteins from a homogeneous mass or a cultured supernatant of said cultured cells, said isolated glycoproteins having the following properties: (a) molecular weight: 12,000-17,000; (b) color reactions: it exhibits a color indicating proteins in the Lowry reaction, exhibits a color indicating peptide bonds and amino acids in the ninhydrin reaction after hydrolysis with hydrochloric acid, and exhibits a color indicating sugars in the phenol-sulfuric acid reaction, the anthrone-sulfuric acid reaction, the indole-sulfuric acid reaction and the tryptophane-sulfuric acid reaction; (c) appearance and solubility: white powder soluble in water, aqueous sodium chloride and phosphate buffer, and sparingly soluble in benzene, hexane and chloroform; (d) sugar content: sugar content is 27-33%, 17-20% of the total sugar being hexoses, 5-7% being hexosamines and 5-6% being sialic acids; (e) isoelectric point: 4.2-7.3; (f) adsorbability: adsorbable on Ulex europeus agglutinin-conjugated Sephadex in 0.01M phosphate buffer (pH 7.2); (g) stability: stable in an aqueous solution of pH 2.0, pH 7.0 or pH 11.0 at 4° C. for 24 hours or longer and in an aqueous solution of pH 7.0 at 60° C. for 3 hours or longer; (h) cytotoxicity: it selectively damages tumor cells without substantially damaging normal cells; and (i) differentiation: induces differentiation of tumor cells.

Claim 11

Dependent Claim 231 words

A process for producing a glycoprotein (CB X1 ) having an anti-tumor effect, comprising: culturing non-established cells or cells of established cell lines of reticulo-endothelial cells, lymphoblasts, leukemia cells or fibroblasts originated from warm-blooded animals, and isolating glycoproteins from a homogeneous mass or a cultured supernatant of said cultured cells, said isolated glycoproteins having the following properties: (a) molecular weight: 70,000-90,000; (b) color reactions: it exhibits a color indicating proteins in the Lowry reaction, exhibits a color indicating peptide bonds and amino acids in the ninhydrin reaction after hydrolysis with hydrochloric acid, and exhibits a color indicating sugars in the phenol-sulfuric acid reaction, the anthrone-sulfuric acid reaction, the indole-sulfuric acid reaction and the tryptophane-sulfuric acid reaction; (c) appearance and solubility: white powder soluble in water, aqueous sodium chloride and phosphate buffer, and sparingly soluble in benzene, hexane and chloroform; (d) sugar content: sugar content is 35-45%, 23-28% of the total sugar being hexoses, 8-11% being hexosamines and 4-6% being sialic acids; (e) isoelectric point: 4.3-6.2; (f) adsorbability: adsorbable on Ulex europeus agglutinin-conjugated Sephadex in 0.01M phosphate buffer (pH 7.2); (g) stability: stable in an aqueous solution of pH 2.0, pH 7.0 or pH 11.0 at 4° C. for 24 hours or longer and in an aqueous solution of pH 7.0 at 60° C. for 3 hours or longer; and (h) cytotoxicity: it selectively damages tumor cells without substantially damaging normal cells.

Claim 12

Dependent Claim 231 words

A process for producing a glycoprotein (CB X1 ) having an anti-tumor effect, comprising: culturing non-established cells or cells of established cell lines of reticulo-endothelial cells, lymphoblasts, leukemia cells or fibroblasts originated from warm-blooded animals, and isolating glycoproteins from a homogeneous mass or a cultured supernatant of said cultured cells, said isolated glycoproteins having the following properties: (a) molecular weight: 70,000-90,000; (b) color reactions: it exhibits a color indicating proteins in the Lowry reaction, exhibits a color indicating peptide bonds and amino acids in the ninhydrin reaction after hydrolysis with hydrochloric acid, and exhibits a color indicating sugars in the phenol-sulfuric acid reaction, the anthrone-sulfuric acid reaction, the indole-sulfuric acid reaction and the tryptophane-sulfuric acid reaction; (c) appearance and solubility: white powder soluble in water, aqueous sodium chloride and phosphate buffer, and sparingly soluble in benzene, hexane and chloroform; (d) sugar content: sugar content is 35-45%, 23-28% of the total sugar being hexoses, 8-11% being hexosamines and 4-6% being sialic acids; (e) isoelectric point: 4.3-6.2; (f) adsorbability: adsorbable on Ulex europeus agglutinin-conjugated Sephadex in 0.01M phosphate buffer (pH 7.2); (g) stability: stable in an aqueous solution of pH 2.0, pH 7.0 or pH 11.0 at 4° C. for 24 hours or longer and in an aqueous solution of pH 7.0 at 60° C. for 3 hours or longer; and (h) cytotoxicity: it selectively damages tumor cells without substantially damaging normal cells.

Claim 13

Dependent Claim 231 words

A process for producing a glycoprotein (CB X2 ) having an anti-tumor effect, comprising: culturing non-established cells or cells of established cell lines of reticulo-endothelial cells, lymphoblasts, leukemia cells or fibroblasts originated from warm-blooded animals, and isolating glycoproteins from a homogeneous mass or a cultured supernatant of said cultured cells, said isolated glycoproteins having the following properties: (a) molecular weight: 40,000-50,000; (b) color reactions: it exhibits a color indicating proteins in the Lowry reaction, exhibits a color indicating peptide bonds and amino acids in the ninhydrin reaction after hydrolysis with hydrochloric acid, and exhibits a color indicating sugars in the phenol-sulfuric acid reaction, the anthrone-sulfuric acid reaction, the indole-sulfuric acid reaction and the tryptophane-sulfuric acid reaction; (c) appearance and solubility: white powder soluble in water, aqueous sodium chloride and phosphate buffer, and sparingly soluble in benzene, hexane and chloroform; (d) sugar content: sugar content is 30-37%, 20-23% of the total sugar being hexoses, 6-8% being hexosamines and 4-6% being sialic acids; (e) isoelectric point: 4.2-7.3; (f) adsorbability: adsorbable on Ulex europeus agglutinin-conjugated Sephadex in 0.01M phosphate buffer (pH 7.2); (g) stability: stable in an aqueous solution of pH 2.0, pH 7.0 or pH 11.0 at 4° C. for 24 hours or longer and in an aqueous solution of pH 7.0 at 60° C. for 3 hours or longer; and (h) cytotoxicity: it selectively damages tumor cells without substantially damaging normal cells.

Claim 14

Dependent Claim 231 words

A process for producing a glycoprotein (CB X2 ) having an anti-tumor effect, comprising: culturing non-established cells or cells of established cell lines of reticulo-endothelial cells, lymphoblasts, leukemia cells or fibroblasts originated from warm-blooded animals, and isolating glycoproteins from a homogeneous mass or a cultured supernatant of said cultured cells, said isolated glycoproteins having the following properties: (a) molecular weight: 40,000-50,000; (b) color reactions: it exhibits a color indicating proteins in the Lowry reaction, exhibits a color indicating peptide bonds and amino acids in the ninhydrin reaction after hydrolysis with hydrochloric acid, and exhibits a color indicating sugars in the phenol-sulfuric acid reaction, the anthrone-sulfuric acid reaction, the indole-sulfuric acid reaction and the tryptophane-sulfuric acid reaction; (c) appearance and solubility: white powder soluble in water, aqueous sodium chloride and phosphate buffer, and sparingly soluble in benzene, hexane and chloroform; (d) sugar content: sugar content is 30-37%, 20-23% of the total sugar being hexoses, 6-8% being hexosamines and 4-6% being sialic acids; (e) isoelectric point: 4.2-7.3; (f) adsorbability: adsorbable on Ulex europeus agglutinin-conjugated Sephadex in 0.01M phosphate buffer (pH 7.2); (g) stability: stable in an aqueous solution of pH 2.0, pH 7.0 or pH 11.0 at 4° C. for 24 hours or longer and in an aqueous solution of pH 7.0 at 60° C. for 3 hours or longer; and (h) cytotoxicity: it selectively damages tumor cells without substantially damaging normal cells.

Claim 15

Dependent Claim 246 words

A process for producing a glycoprotein (CB X3 ) having an anti-tumor effect, comprising: culturing non-established cells or cells of established cell lines of reticulo-endothelial cells, lymphoblasts, leukemia cells or fibroblasts originated from warm-blooded animals, and isolating glycoproteins from a homogeneous mass or a cultured supernatant of said cultured cells, said isolated glycoproteins having the following properties: (a) molecular weight: 7,000-9,000; (b) color reactions: it exhibits a color indicating proteins in the Lowry reaction, exhibits a color indicating peptide bonds and amino acids in the ninhydrin reaction after hydrolysis with hydrochloric acid, and exhibits a color indicating sugars in the phenol-sulfuric acid reaction, the anthrone-sulfuric acid reaction, the indole-sulfuric acid reaction and the tryptophane-sulfuric acid reaction; (c) appearance and solubility: white powder soluble in water, aqueous sodium chloride and phosphate buffer, and sparingly soluble in benzene, hexane and chloroform; (d) sugar content: sugar content is 8-15%, 6-10% of the total sugar being hexoses, 1-2% being hexosamines and 1-3% being sialic acids; (e) adsorbability: adsorbable on carboxymethylcellulose in an ion exchange chromatography in 0.05M phosphate buffer (pH 6.4) using carboxymethylcellulose; (f) stability: stable in an aqueous solution of pH 2.0, pH 7.0 or pH 11.0 at 4° C. for 24 hours or longer and in an aqueous solution of pH 7.0 at 60° C. for 3 hours or longer; (g) cytotoxicity: it selectively damages tumor cells without substantially damaging normal cells; and (h) the amino acid sequence of the N terminal of its protein portion is Alanine-Alanine.

Claim 16

Dependent Claim 246 words

A process for producing a glycoprotein (CB X3 ) having an anti-tumor effect, comprising: culturing non-established cells or cells of established cell lines of reticulo-endothelial cells, lymphoblasts, leukemia cells or fibroblasts originated from warm-blooded animals, and isolating glycoproteins from a homogeneous mass or a cultured supernatant of said cultured cells, said isolated glycoproteins having the following properties: (a) molecular weight: 7,000-9,000; (b) color reactions: it exhibits a color indicating proteins in the Lowry reaction, exhibits a color indicating peptide bonds and amino acids in the ninhydrin reaction after hydrolysis with hydrochloric acid, and exhibits a color indicating sugars in the phenol-sulfuric acid reaction, the anthrone-sulfuric acid reaction, the indole-sulfuric acid reaction and the tryptophane-sulfuric acid reaction; (c) appearance and solubility: white powder soluble in water, aqueous sodium chloride and phosphate buffer, and sparingly soluble in benzene, hexane and chloroform; (d) sugar content: sugar content is 8-15%, 6-10% of the total sugar being hexoses, 1-2% being hexosamines and 1-3% being sialic acids; (e) adsorbability: adsorbable on carboxymethylcellulose in an ion exchange chromatography in 0.05M phosphate buffer (pH 6.4) using carboxymethylcellulose; (f) stability: stable in an aqueous solution of pH 2.0, pH 7.0 or pH 11.0 at 4° C. for 24 hours or longer and in an aqueous solution of pH 7.0 at 60° C. for 3 hours or longer; (g) cytotoxicity: it selectively damages tumor cells without substantially damaging normal cells; and (h) the amino acid sequence of the N terminal of its protein portion is Alanine-Alanine.

Claim 17

Dependent Claim 20 words

A process according to claim 5, wherein the source cells are exposed to the action of one or more inducers.

Claim 18

Dependent Claim 27 words

A process for producing a glycoprotein having an anti-tumor effect as claimed in claim 6, wherein said cultured cells are exposed to the action of an inducer.

Claim 19

Dependent Claim 27 words

A process for producing a glycoprotein having an anti-tumor effect as claimed in claim 7, wherein said cultured cells are exposed to the action of an inducer.

Claim 20

Dependent Claim 27 words

A process for producing a glycoprotein having an anti-tumor effect as claimed in claim 8, wherein said cultured cells are exposed to the action of an inducer.

Claim 21

Dependent Claim 50 words

A process for producing a glycoprotein having an anti-tumor effect as claimed in claim 9, wherein the inducer is selected from the group consisting of lectins such as phytohemagglutinin, concanavalin A, pokeweed mitogen, lypopolysaccharides, polysaccharides such as phosphomannan, dextran phosphate, endotoxins, microbial cell components, bacteria, viruses, nucleic acids and polynucleotides.

Claim 22

Dependent Claim 50 words

A process for producing a glycoprotein having an anti-tumor effect as claimed in claim 10, wherein the inducer is selected from the group consisting of lectins such as phytohemagglutinin, concanavalin A, pokeweed mitogen, lypopolysaccharides, polysaccharides such as phosphomannan, dextran phosphate, endotoxins, microbial cell components, bacteria, viruses, nucleic acids and polynucleotides.

Claim 23

Dependent Claim 50 words

A process for producing a glycoprotein having an anti-tumor effect as claimed in claim 11, wherein the inducer is selected from the group consisting of lectins such as phytohemagglutinin, concanavalin A, pokeweed mitogen, lypopolysaccharides, polysaccharides such as phosphomannan, dextran phosphate, endotoxins, microbial cell components, bacteria, viruses, nucleic acids and polynucleotides.

Claim 24

Dependent Claim 50 words

A process for producing a glycoprotein having an anti-tumor effect as claimed in claim 12, wherein the inducer is selected from the group consisting of lectins such as phytohemagglutinin, concanavalin A, pokeweed mitogen, lypopolysaccharides, polysaccharides such as phosphomannan, dextran phosphate, endotoxins, microbial cell components, bacteria, viruses, nucleic acids and polynucleotides.

Claim 25

Dependent Claim 40 words

A process according to claim 5, wherein the established cell lines are selected from the group consisting of BALL-1, TALL-1, NALL-1, Namalwa, M-7002, B-7101, Flow 7000, JBL, EBV-Sa, EBV-Wa, EBV-HO, BALM2 and CCRF-SB, all of which are of human origin.

Claim 26

Dependent Claim 52 words

A process for producing a glycoprotein having an anti-tumor effect as claimed in claim 6, wherein the cells of the established cell lines are selected from the group consisting of BALL-1, TALL-1, NALL-1, Namalwa, M-7002, M-7101, Flow 7000, JBL, EBV-Sa, EBV-Wa, EBV-HO, BALM2 and CCRF-SB, all of which are of human origin.

Claim 27

Dependent Claim 52 words

A process for producing a glycoprotein having an anti-tumor effect as claimed in claim 7, wherein the cells of the established cell lines are selected from the group consisting of BALL-1, TALL-1, NALL-1, Namalwa, M-7002, B-7101, Flow 7000, JBL, EBV-Sa, EBV-Wa, EBV-HO, BALM2 and CCRF-SB, all of which are of human origin.

Claim 28

Dependent Claim 52 words

A process for producing a glycoprotein having an anti-tumor effect as claimed in claim 8, wherein the cells of the established cell lines are selected from the group consisting of BALL-1, TALL-1, NALL-1, Namalwa, M-7002, B-7101, Flow 7000, JBL, EBV-Sa, EBV-Wa, EBV-HO, BALM2 and CCRF-SB, all of which are of human origin.

Claim 29

Dependent Claim 48 words

A process according to claim 5, wherein the established cell lines are selected from the group consisting of mouse BALB/C 3T3, mouse leukemia cells L1210, P388, mouse melanoma clone M-3, rat tumor LLC-WRC 256, and hamster melanoma RPMI 1846, all of which are of non-human warm-blooded animal origin.

Claim 30

Dependent Claim 60 words

A process for producing a glycoprotein having an anti-tumor effect as claimed in claim 6, wherein the cells of the established cell lines are selected from the group consisting of mouse BALB/C 3T3, mouse leukemia cells L1210, P388, mouse melanoma clone M-3, rat tumor LLC-WRC 256, and hamster melanoma RPMI 1846, all of which are of non-human warm-blooded animal origin.

Claim 31

Dependent Claim 60 words

A process for producing a glycoprotein having an anti-tumor effect as claimed in claim 7, wherein the cells of the established cell lines are selected from the group consisting of mouse BALB/C 3T3, mouse leukemia cells L1210, P388, mouse melanoma clone M-3, rat tumor LLC-WRC 256, and hamster melanoma RPMI 1846, all of which are of non-human warm-blooded animal origin.

Claim 32

Dependent Claim 60 words

A process for producing a glycoprotein having an anti-tumor effect as claimed in claim 8, wherein the cells of the established cell lines are selected from the group consisting of mouse BALB/C 3T3, mouse leukemia cells L1210, P388, mouse melanoma clone M-3, rat tumor LLC-WRC 256, and hamster melanoma RPMI 1846, all of which are of non-human warm-blooded animal origin.

Claim 33

Dependent Claim 27 words

A process according to claim 5, wherein said cells are cultured in the body of a non-human animal and grown therein prior to isolation of said glycoproteins.

Claim 34

Dependent Claim 27 words

A process according to claim 6, wherein said cells are cultured in the body of a non-human animal and grown therein prior to isolation of said glycoproteins.

Claim 35

Dependent Claim 27 words

A process according to claim 7, wherein said cells are cultured in the body of a non-human animal and grown therein prior to isolation of said glycoproteins.

Claim 36

Dependent Claim 26 words

A process according to claim 8, wherein said cells cultured in the body of a non-human animal and grown therein prior to isolation of said glycoproteins.

Claim 37

Dependent Claim 19 words

A process according to claim 5, wherein said cells are cultured in vitro prior to isolation of said glycoproteins.

Claim 38

Dependent Claim 19 words

A process according to claim 6, wherein said cells are cultured in vitro prior to isolation of siad glycoproteins.

Claim 39

Dependent Claim 19 words

A process according to claim 7, wherein said cells are cultured in vitro prior to isolation of said glycoproteins.

Claim 40

Dependent Claim 19 words

A process according to claim 8, wherein said cells are cultured in vitro prior to isolation of said glycoproteins.

Claim 41

Dependent Claim 23 words

A process according to claim 5, wherein said cells are cultured in a diffusion chamber through which said animal body fluid medium circulates.

Claim 42

Dependent Claim 21 words

A process according to claim 33, wherein said diffusion chamber is inside an animal during the growth of said cultured cells.

Claim 43

Dependent Claim 23 words

A process according to claim 6, wherein said cells are cultured in a diffusion chamber through which said animal body fluid medium circulates.

Claim 44

Dependent Claim 21 words

A process according to claim 35, wherein said diffusion chamber is inside an animal during the growth of said cultured cells.

Claim 45

Dependent Claim 23 words

A process according to claim 7, wherein said cells are cultured in a diffusion chamber through which said animal body fluid medium circulates.

Claim 46

Dependent Claim 21 words

A process according to claim 37, wherein said diffusion chamber is inside an animal during the growth of said cultured cells.

Claim 47

Dependent Claim 23 words

A process according to claim 8, wherein said cells are cultured in a diffusion chamber through which said animal body fluid medium circulates.

Claim 48

Dependent Claim 21 words

A process according to claim 39, wherein said diffusion chamber is inside an animal during the growth of said cultured cells.

Claim 49

Dependent Claim 21 words

A process according to claim 25, wherein said cultured cells are further cultured in vitro after being grown in said animal.

Claim 50

Dependent Claim 21 words

A process according to claim 26, wherein said cultured cells are further cultured in vitro after being grown in said animal.

Claim 51

Dependent Claim 21 words

A process according to claim 27, wherein said cultured cells are further cultured in vitro after being grown in said animal.

Claim 52

Dependent Claim 21 words

A process according to claim 28, wherein said cultured cells are further cultured in vitro after being grown in said animal.

Claim 53

Dependent Claim 22 words

A process according to claim 33, wherein said cultured cells are further cultured in vitro after being grown in said diffusion chamber.

Claim 54

Dependent Claim 22 words

A process according to claim 35, wherein said cultured cells are further cultured in vitro after being grown in said diffusion chamber.

Claim 55

Dependent Claim 22 words

A process according to claim 37, wherein said cultured cells are further cultured in vitro after being grown in said diffusion chamber.

Claim 56

Dependent Claim 22 words

A process according to claim 39, wherein said cultured cells are further cultured in vitro after being grown in said diffusion chamber.

Claim 57

Dependent Claim 35 words

A process according to claim 25, wherein the possibility of immunological reactions of the animals to which cells are transplanted is minimized by exposing the animals to X-rays or by injecting them with immunosuppressive agents.

Claim 58

Dependent Claim 35 words

A process according to claim 26, wherein the possibility of immunological reactions of the animals to which cells are transplanted is minimized by exposing the animals to X-rays or by injecting them with immunosuppressive agents.

Claim 59

Dependent Claim 35 words

A process according to claim 27, wherein the possibility of immunological reactions of the animals to which cells are transplanted is minimized by exposing the animals to X-rays or by injecting them with immunosuppressive agents.

Claim 60

Dependent Claim 35 words

A process according to claim 28, wherein the possibility of immunological reactions of the animals to which cells are transplanted is minimized by exposing the animals to X-rays or by injecting them with immunosuppressive agents.

Claim 61

Dependent Claim 17 words

A process according to claim 5, wherein the animals to which cells are transplanted are nude mice.

Claim 62

Dependent Claim 17 words

A process according to claim 6, wherein the animals to which cells are transplanted are nude mice.

Claim 63

Dependent Claim 17 words

A process according to claim 7, wherein the animals to which cells are transplanted are nude mice.

Claim 64

Dependent Claim 17 words

A process according to claim 8, wherein the animals to which cells are transplanted are nude mice.

Claim 65

Dependent Claim 21 words

A process according to claim 5, wherein the non-established cells are selected from the group consisting of macrophages and human lymphocytes.

Claim 66

Dependent Claim 20 words

A process according to claim 6, wherein the non-established cells are selected from the group consisting of lymphocytes and macrophages.

Claim 67

Dependent Claim 29 words

A process for producing a glycoprotein having an anti-tumor effect as claimed in claim 7, wherein the non-established cells are selected from the group consisting of macrophages and lymphocytes.

Claim 68

Dependent Claim 29 words

A process for producing a glycoprotein having an anti-tumor effect as claimed in claim 8, wherein the non-established cells are selected from the group consisting of macrophages and lymphocytes.

Annuity Stage	Window Opens	Payment Deadline	Surcharge Period Until	Estimated Fee	Status
3.5 Years (Year 4)	1987-11-06	1988-05-07	1988-11-06	$2000.00	Window Closed / Lapsed
7.5 Years (Year 8)	1991-11-06	1992-05-07	1992-11-06	$3760.00	Window Closed / Lapsed
11.5 Years (Year 12)	1995-11-06	1996-05-07	1996-11-06	$7700.00	Window Closed / Lapsed

Glycoproteins and processes for their production

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