US5122450A
Expired / Lapsed
US
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Biochemical reagent
Assignee: RESEARCH Corp Ltd 91 WATERLOO ROAD LONDON SE1 8XP UNITED KINGDM A CORP OF UNITED KINGDOM
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Filing: 1986-10-24
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Granted: 1992-06-16
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Expires: 2006-10-24
Abstract
Abstract A biochemical reagent comprises an oligosaccharide, preferably one which has been liberated from an immunogenic glycoprotein or proteoglycan, which is immobilized on a carrier via an intermediate spacer molecule such as a lipid. The lipid molecule should preferably have at least two long lipid tails so that the oligosaccharide is held in spaced relationship to the carrier where is exhibits antibody-binding ability which is almost indistinguishable from that of the original glycoprotein or proteoglycan. The reagent has its application in biochemical testing of oligosaccharides and systems which bind to them.
Inventors
- 👤 Ten Feizi
- 👤 Ping W. Tang
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-06 14:07
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54313; G01N33/54353; Y; Y10; Y10S; Y10S435/00; Y10S435/81; Y10S435/814; Y10S436/00; Y10S436/807; Y10S436/808; Y10S436/809; Y10S436/823; Y10S436/824; Y10S530/00; Y10S530/81; Y10S530/811; Y10S530/812; Y10S530/815; Y10S530/816
Claims Analysis
Total Claims: 62
Claim 1
Independent Claim
69 words
An immobilized biochemical reagent comprising (a) a solid immobilizing carrier having bound thereon (b) a conjugate which is a reaction product of (1) an oligosaccharide and (2) a lipid spacer molecule having at least two hydrophobic chains, whereby the oligosaccharide is held in spaced relationship to said solid immobilizing carrier by said lipid spacer molecule and is presented in antigenically active steric configuration for binding on encountering an antibody.
Claim 2
Dependent Claim
69 words
An immobilized biochemical reagent comprising (a) a solid immobilizing carrier having bound thereon (b) a conjugate which is a reaction product of (1) an oligosaccharide and (2) a lipid spacer molecule having at least two hydrophobic chains, whereby the oligosaccharide is held in spaced relationship to said solid immobilizing carrier by said lipid spacer molecule and is presented in antigenically active steric configuration for binding on encountering an antibody.
Claim 3
Dependent Claim
22 words
The reagent as claimed in claim 1, wherein said hydrophobic chains comprise linear alkyl groups containing from 5 to 50 carbon atoms.
Claim 4
Dependent Claim
98 words
The reagent as claimed in claim 1, in which said oligosaccharide and said lipid form a neoglycolipid in which the neoglycolipid has the general formula: ##STR5## wherein R 1 represents a carbohydrate group derived from said oligosaccharide, R 2 represents an alkyl or alkoyl group, R 3 represents an alkyl or alkoyl group, R 4 represents an alkyl or alkoyl group, or hydrogen, R 2 , R 3 and R 4 being the same or different groups, X represents a linking group selected from the group consisting of: ##STR6## and z is zero or 1, 2 or 3.
Claim 5
Dependent Claim
39 words
The reagent as claimed in claim 3 in which the group X is a residue of a compound selected from the group consisting of phosphatidyl ethanolamine dipalmitoyl (PPEADP), L-alpha-phosphatidyl-L-serine, L-alpha-phosphatidyl-ethanolamine (PPEA) dilauroyl, PPEA dimyristoyl, PPEA distearoyl, and PPEA beta-oleoyl-gamma-palmitoyl.
Claim 6
Dependent Claim
18 words
The reagent as claimed in claim 4 in which the oligosaccharide is derived from a glycoprotein or proteoglycan.
Claim 7
Dependent Claim
30 words
The reagent as claimed in claim 4 in which the solid immobilizing carrier is selected from the group consisting of a chromatographic support material, and a plate of plastics material.
Claim 8
Dependent Claim
15 words
The reagent as claimed in claim 6 in which the plastics material is polyvinyl chloride.
Claim 9
Dependent Claim
16 words
The reagent as claimed in claim 6 in which the chromatographic support material is silica gel.
Claim 10
Dependent Claim
43 words
The reagent as claimed in claim 3, wherein each of R 2 , R 3 and R 4 independently of the others represents a linear C 5 to C 50 alkyl group or a linear (C 5 to C 50 alkyl) carbonyl group.
Claim 11
Dependent Claim
43 words
The reagent as claimed in claim 9, wherein each of R 2 , R 3 and R 4 independently of the others represents a linear C 10 to C 25 alkyl group or a linear (C 10 to C 25 alkyl) carbonyl group.
Claim 12
Dependent Claim
20 words
The reagent as claimed in claim 1 or 3 in which the oligosaccharide is derived from a glycoprotein or proteoglycan.
Claim 13
Dependent Claim
32 words
The reagent as claimed in claim 1 or 3 in which the solid immobilizing carrier is selected from the group consisting of a chromatographic support material, and a plate of plastics material.
Claim 14
Dependent Claim
62 words
A biochemical assay kit comprising (1) an immobilized biochemical reagent as claimed in any one of claim 1, 3, 4, 5 or 6 in which said solid immobilizing carrier is a test plate and (2) at least one further reagent selected from a solution of a neoglycolipid of which the glyco group is the said oligosaccharide and a solution of liposome-forming components.
Claim 15
Dependent Claim
54 words
A biochemical assay kit comprising (1) an immobilized biochemical reagent as claimed in claim 11 in which said solid immobilizing carrier is a test plate, (2) a reagent comprising a solution of a neoglycolipid of which the glyco group is the said oligosaccharide and (3) a further reagent comprising a solution of liposome-forming components.
Claim 16
Dependent Claim
54 words
A biochemical assay kit comprising (1) an immobilized biochemical reagent as claimed in claim 12 in which said solid immobilizing carrier is a test plate, (2) a reagent comprising a solution of a neoglycolipid of which the glyco group is the said oligosaccharide and (3) a further reagent comprising a solution of liposome-forming components.
Claim 17
Independent Claim
45 words
A method of preparing an immobilized biochemical reagent, comprising reacting a reducing oligosaccharide containing a reactive aldehyde group and an amine derivative of a lipid having at least two hydrophobic chains to produce a neoglycolipid and bonding the reaction product to a solid immobilizing carrier.
Claim 18
Independent Claim
45 words
A method of preparing an immobilized biochemical reagent, comprising reacting a reducing oligosaccharide containing a reactive aldehyde group and an amine derivative of a lipid having at least two hydrophobic chains to produce a neoglycolipid and bonding the reaction product to a solid immobilizing carrier.
Claim 19
Dependent Claim
22 words
The method as claimed in claim 16, wherein said hydrophobic chains comprise linear alkyl groups containing from 5 to 50 carbon atoms.
Claim 20
Dependent Claim
32 words
The method as claimed in claim 16, wherein said hydrophobic chains are selected from linear C 10 to C 25 alkyl groups and linear (C 10 to C 25 alkyl) carbonyl groups.
Claim 21
Dependent Claim
26 words
The method of preparing an immobilized biochemical reagent as claimed in claim 16 wherein said oligosaccharide is an oligosaccharide liberated from an immunogenic glycoprotein or proteoglycan.
Claim 22
Dependent Claim
28 words
The method of preparing an immobilized biochemical reagent as claim in claim 19 wherein said oligosaccharide is liberated by contacting said glycoprotein or said proteoglycan with an enzyme.
Claim 23
Dependent Claim
42 words
The method of preparing an immobilized biochemical reagent as claimed in claim 19 wherein said oligosaccharide is liberated by contacting said glycoprotein or said proteoglycan with alkaline borohydride, followed by oxidizing said liberated oligosaccharide with periodate to form an aldehyde group thereon.
Claim 24
Dependent Claim
35 words
The method of preparing an immobilized biochemical reagent as claimed in claim 19 wherein said oligosaccharide is liberated by contacting said glycoprotein or said proteoglycan with hydrazine, followed by re-N-acylation to form a re-N-acylated oligosaccharide.
Claim 25
Dependent Claim
35 words
The method of preparing an immobilized biochemical reagent as claimed in claim 22 wherein said amine derivative of said lipid and said re-N-acylated oligosaccharide are reacted in the presence of cyanoborohydride to form a neoglycolipid.
Claim 26
Dependent Claim
49 words
The method as claimed in claim 22, wherein said re-N-acylated oligosaccharide is reacted with sodium borohydride to cleave the terminal saccharide ring structure with formation of a methylol group and then with periodate to form a reducing aldehyde group for reaction with an amine group on the lipid derivative.
Claim 27
Independent Claim
113 words
A method of conducting a biochemical assay, comprising providing an immobilized biochemical reagent comprising (a) a solid immobilizing carrier having bound thereon (b) a conjugate which is a reaction product of (1) an oligosaccharide and (2) a lipid spacer molecule having at least two hydrocarbon chains, whereby the oligosaccharide is held in spaced relationship to said solid immobilizing carrier by said lipid spacer molecule and is presented in antigenically active steric configuration for binding on encountering an antibody; (3) contacting said conjugate with an antibody to the oligosaccharide to form an antibody-conjugate complex whereby the antibody binds to the oligosaccharide of said conjugate; and (4) detecting said complex with an analytically detectable reagent.
Claim 28
Independent Claim
113 words
A method of conducting a biochemical assay, comprising providing an immobilized biochemical reagent comprising (a) a solid immobilizing carrier having bound thereon (b) a conjugate which is a reaction product of (1) an oligosaccharide and (2) a lipid spacer molecule having at least two hydrocarbon chains, whereby the oligosaccharide is held in spaced relationship to said solid immobilizing carrier by said lipid spacer molecule and is presented in antigenically active steric configuration for binding on encountering an antibody; (3) contacting said conjugate with an antibody to the oligosaccharide to form an antibody-conjugate complex whereby the antibody binds to the oligosaccharide of said conjugate; and (4) detecting said complex with an analytically detectable reagent.
Claim 29
Dependent Claim
22 words
The method as claimed in claim 25, wherein said hydrophobic chains comprise linear alkyl groups containing from 5 to 50 carbon atoms.
Claim 30
Dependent Claim
32 words
The method as claimed in claim 25, wherein said hydrophobic chains are selected from linear C 10 to C 25 alkyl groups and linear (C 1 to C 25 alkyl) carbonyl groups.
Claim 31
Dependent Claim
41 words
The method of conducting a biochemical assay as claimed in claim 25, which includes an inhibition assay step comprising contacting said complex with a second biochemical reagent comprising a conjugate of an oligosaccharide with a solid phase or a liposome carrier.
Claim 32
Dependent Claim
25 words
The method of conducting a biochemical assay as claimed in claim 25, wherein the analytically detectable reagent is a radiolabelled antibody to the bound antibody.
Claim 33
Dependent Claim
17 words
The method as claimed in claim 29, wherein detection of said radiolabelled antibody is effected by autoradiography.
Claim 34
Dependent Claim
84 words
An immobilized biochemical reagent comprising (a) a solid immobilizing carrier having bound therein (b) a conjugate which is a reaction product of (1) an oligosaccharide selected from the group consisting of monosaccharides and oligosaccharides containing up to 12 saccharide residues, and (2) a lipid spacer molecule having at least two hydrophobic chains, whereby the oligosaccharide is held in spaced relationship to said solid immobilizing carrier by said lipid spacer molecule and is presented in antigenically active steric configuration for binding on encountering an antibody.
Claim 35
Dependent Claim
84 words
An immobilized biochemical reagent comprising (a) a solid immobilizing carrier having bound therein (b) a conjugate which is a reaction product of (1) an oligosaccharide selected from the group consisting of monosaccharides and oligosaccharides containing up to 12 saccharide residues, and (2) a lipid spacer molecule having at least two hydrophobic chains, whereby the oligosaccharide is held in spaced relationship to said solid immobilizing carrier by said lipid spacer molecule and is presented in antigenically active steric configuration for binding on encountering an antibody.
Claim 36
Dependent Claim
22 words
The reagent as claimed in claim 31, wherein said hydrophobic chains comprise linear alkyl groups containing from 5 to 50 carbon atoms.
Claim 37
Dependent Claim
18 words
The reagent as claimed in claim 31 in which the oligosaccharide is derived from a glycoprotein or proteoglycan.
Claim 38
Dependent Claim
100 words
The reagent as claim in claim 31 in which said oligosaccharide and said lipid form a neoglycolipid in which the neoglycolipid has the general formula: ##STR7## wherein R 1 represents a carbohydrate group derived from said oligosaccharide, R 2 represents an alkyl or alkoyl group, R 3 represents an alkyl or alkoyl group, R 4 represents an alkyl or alkoyl group, or hydrogen, R 2 , R 3 , and R 4 being the same or different groups, and X represents a linking group selected from the group consisting of: ##STR8## and z is zero or 1, 2 or 3.
Claim 39
Dependent Claim
43 words
The reagent as claimed in claim 34, wherein each of R 2 , R 3 and R 4 independently of the others represents a linear C 5 to C 50 alkyl group or a linear (C 5 to C 50 alkyl) carbonyl group.
Claim 40
Dependent Claim
43 words
The reagent as claimed in claim 35, wherein each of R 2 , R 3 and R 4 independently of the others represents a linear C 10 to C 25 alkyl group or a linear (C 10 to C 25 alkyl) carbonyl group.
Claim 41
Dependent Claim
39 words
The reagent as claimed in claim 34, in which the group X is a residue of a compound selected from the group consisting of phosphatidyl ethanolamine dipalmitoyl (PPEADP), L-alpha-phosphatidyl-L-serine, L-alpha-phosphatidyl-ethanolamine (PPEA) dilauroyl, PPEA dimyristoyl, PPEA distearoyl, and PPEA beta-oleoyl-gamma-palmitoyl.
Claim 42
Dependent Claim
30 words
The reagent as claim in claim 37 in which the solid immobilizing carrier is selected from the group consisting of a chromatographic support material and a plate of plastics material.
Claim 43
Dependent Claim
15 words
The reagent as claimed in claim 38 in which the plastics material is polyvinyl chloride.
Claim 44
Dependent Claim
16 words
The reagent as claimed in claim 38 in which the chromatographic support material is silica gel.
Claim 45
Dependent Claim
20 words
The reagent as claimed in claim 31 or 34 in which the oligosaccharide is derived from a glycoprotein or proteoglycan.
Claim 46
Dependent Claim
32 words
The reagent as claimed in claim 31 or 34, in which the solid immobilizing carrier is selected from the group consisting of a chromatographic support material and a plate of plastics material.
Claim 47
Dependent Claim
52 words
A biochemical assay kit comprising (1) an immobilized biochemical reagent as claimed in claim 41, in which said solid immobilizing carrier is a test plate (2) a reagent comprising solution of neoglycolipid of which the glyco group is the said oligosaccharide and (3) a further reagent comprising a solution of liposome-forming components.
Claim 48
Dependent Claim
53 words
A biochemical assay kit comprising (1) an immobilized biochemical reagent as claimed in claim 42, in which said solid immobilizing carrier is a test plate, (2) a reagent comprising solution of a neoglycolipid of which the glyco groups is the said oligosaccharide and (3) a further reagent comprising a solution of liposome-forming components.
Claim 49
Dependent Claim
61 words
A biochemical assay kit comprising (1) an immobilized biochemical reagent as claimed in any one of claims 31, 34, 37, 33 or 38, in which said solid immobilizing carrier is a test plate and (2) at least one further reagent selected from a solution of neoglycolipid of which the glyco group is the said oligosaccharide and a solution of liposome-forming components.
Claim 50
Independent Claim
79 words
A method of preparing an immobilized biochemical reagent comprising reacting a reducing oligosaccharide, which contains an aldehyde group and which is selected from the group consisting of monosaccharides and oligosaccharides containing up to 12 saccharide residues, and an amine derivative of a lipid having at least two hydrophobic chains to produce a neoglycolipid, and bonding the reaction product to a solid immobilizing carrier whereby the oligosaccharide is held in spaced relationship to said solid immobilizing carrier by said lipid.
Claim 51
Independent Claim
79 words
A method of preparing an immobilized biochemical reagent comprising reacting a reducing oligosaccharide, which contains an aldehyde group and which is selected from the group consisting of monosaccharides and oligosaccharides containing up to 12 saccharide residues, and an amine derivative of a lipid having at least two hydrophobic chains to produce a neoglycolipid, and bonding the reaction product to a solid immobilizing carrier whereby the oligosaccharide is held in spaced relationship to said solid immobilizing carrier by said lipid.
Claim 52
Dependent Claim
26 words
The method of preparing an immobilized biochemical reagent as claimed in claim 46 wherein said oligosaccharide is an oligosaccharide liberated from an immunogenic glycoprotein or proteoglycan.
Claim 53
Dependent Claim
28 words
The method of preparing an immobilized biochemical reagent as claimed in claim 47 wherein said oligosaccharide is liberated by contacting said glycoprotein or said proteoglycan with an enzyme.
Claim 54
Dependent Claim
42 words
The method of preparing an immobilized biochemical reagent as claimed in claim 47 wherein said oligosaccharide is liberated by contacting said glycoprotein or said proteoglycan with alkaline borohydride, followed by oxidizing said liberated oligosaccharide with periodate to form an aldehyde group thereon.
Claim 55
Dependent Claim
35 words
The method of preparing an immobilized biochemical reagent as claimed in claim 47, wherein said oligosaccharide is liberated by contacting said glycoprotein or said proteoglycan with hydrazine, followed by re-N-acylation to form a re-N-acylated oligosaccharide.
Claim 56
Dependent Claim
55 words
The method of preparing an immobilized biochemical reagent as claimed in claim 50, wherein said re-N-acylated oligosaccharide is reacted with sodium borohydride to cleave the terminal saccharide ring structure with formation of a methylol group and then with periodate to form a reducing aldehyde group for reaction with an amine group on the lipid derivative.
Claim 57
Dependent Claim
35 words
The method of preparing an immobilized biochemical reagent as claimed in claim 50 wherein said amine derivative of said lipid and said re-N-acylated oligosaccharide are reacted in the presence of cyanoborohydride to form a neoglycolipid.
Claim 58
Independent Claim
126 words
A method of conducting a biochemical assay, comprising providing an immobilized biochemical reagent comprising (a) a solid immobilizing carrier having bound thereon (b) a conjugate which is a reaction product of (1) an oligosaccharide selected from the group consisting of monosaccharides and oligosaccharides containing up to 12 saccharide residues, and (2) a lipid spacer molecule having at least two hydrophobic chains, whereby the oligosaccharide is held in spaced relationship to said solid immobilizing carrier by said lipid spacer molecule and is presented in antigenically active steric configuration for binding on encountering an antibody; contacting said conjugate with an antibody to the oligosaccharide to form an antibody-conjugate complex whereby the antibody binds to the oligosaccharide of said conjugate; and detecting said complex with an analytically detectable reagent.
Claim 59
Independent Claim
126 words
A method of conducting a biochemical assay, comprising providing an immobilized biochemical reagent comprising (a) a solid immobilizing carrier having bound thereon (b) a conjugate which is a reaction product of (1) an oligosaccharide selected from the group consisting of monosaccharides and oligosaccharides containing up to 12 saccharide residues, and (2) a lipid spacer molecule having at least two hydrophobic chains, whereby the oligosaccharide is held in spaced relationship to said solid immobilizing carrier by said lipid spacer molecule and is presented in antigenically active steric configuration for binding on encountering an antibody; contacting said conjugate with an antibody to the oligosaccharide to form an antibody-conjugate complex whereby the antibody binds to the oligosaccharide of said conjugate; and detecting said complex with an analytically detectable reagent.
Claim 60
Dependent Claim
41 words
The method of conducting a biochemical assay as claimed in claim 53 which includes an inhibition assay step comprising contacting said complex with a second biochemical reagent comprising a conjugate of an oligosaccharide with a solid phase or a liposome carrier.
Claim 61
Dependent Claim
25 words
The method of conducting a biochemical assay as claimed in claim 53, wherein the analytically detectable reagent is a radiolabelled antibody to the bound antibody.
Claim 62
Dependent Claim
22 words
The method of conducting a biochemical assay as claimed in claim 53, wherein detection of said radiolabelled antibody is effected by autoradiography.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1995-06-16 | 1995-12-16 | 1996-06-16 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1999-06-16 | 1999-12-16 | 2000-06-16 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2003-06-16 | 2003-12-16 | 2004-06-16 | $7700.00 | Window Closed / Lapsed |