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Formation of colloidal metal dispersions using aminodextrans as reductants and protective agents

Assignee: Beckman Coulter Inc
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Filing: 1992-01-29
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Granted: 1993-09-28
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Expires: 2012-01-29
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Abstract

Abstract The invention related generally to the preparation of colloidal metal(O) particles having a crosslinked aminodextran coating with pendent amine groups attached thereto. The aminodextran acts as both a reductant for reducing metal ions to metal(O) particles and as the protective agent which coats the metal(O) particles thus formed. After stabilizing the aminodextran coating by use of a crosslinking agent, the coated particles can be used to covalently bind proteins. The resulting protein containing colloidal particles can be used as markers in optical and electron microscopy, in immunological and biological assays, and possibly as therapeutic agents.

Inventors

  • 👤 Olavi Siiman
  • 👤 Alexander Burshteyn

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-11-07 10:06
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/551; G01N33/553; A; A61; A61K; A61K47/00; A61K47/50; A61K47/69; A61K47/6921; A61K47/6923; A61K47/6927; A61K47/6929; G01N33/544; G01N33/548; Y; Y10; Y10T; Y10T428/00; Y10T428/29; Y10T428/2982; Y10T428/2989; Y10T428/2991; Y10T428/2998

Claims Analysis

Total Claims: 41
Claim 1
Independent Claim 112 words
A method for preparing colloidal metal(O) particles from a solution of a metal salt and coating said particles with a polymeric organic compound which also is used as a reductant for said metal salt, said method comprising: (a) heating a solution containing an amine derivatized, oxidizable sugar containing polysaccharide and a metal salt capable of being reduced by said oxidizable sugar containing polysaccharide for a time and at a temperature sufficient to reduce said metal salt to colloidal metal particles and simultaneously coating said particles with said polysaccharide; and (b) stabilizing the amine derivatized polysaccharide coating on said particles by means of a bifunctional crosslinking agent, a diamine and a reducing agent.
Claim 2
Independent Claim 112 words
A method for preparing colloidal metal(O) particles from a solution of a metal salt and coating said particles with a polymeric organic compound which also is used as a reductant for said metal salt, said method comprising: (a) heating a solution containing an amine derivatized, oxidizable sugar containing polysaccharide and a metal salt capable of being reduced by said oxidizable sugar containing polysaccharide for a time and at a temperature sufficient to reduce said metal salt to colloidal metal particles and simultaneously coating said particles with said polysaccharide; and (b) stabilizing the amine derivatized polysaccharide coating on said particles by means of a bifunctional crosslinking agent, a diamine and a reducing agent.
Claim 3
Dependent Claim 13 words
The method of claim 1 wherein said amine derivatized polysaccharide is an aminodextran.
Claim 4
Dependent Claim 18 words
The method of claim 1 wherein said metal salt has a reduction potential of +0.7 volts or higher.
Claim 5
Dependent Claim 31 words
The method of claim 3 wherein said metal salt is selected from the group consisting of gold, silver, platinum, palladium, rhodium and iridium salts capable of being reduced by said polysaccharide.
Claim 6
Dependent Claim 15 words
The method of claim 4 wherein said metal salt is a gold or silver salt.
Claim 7
Independent Claim 179 words
A method of preparing colloidal metal(O) particles coated with an amine derivatized, oxidizable sugar containing polysaccharide, said method comprising: (a) heating an aqueous solution of a sugar reducible metal salt and adding to said solution an aqueous solution of an amine derivatized polysaccharide having at least one reducing sugar component; (b) heating and mixing the solution resulting from step (a) for about 10-30 minutes at a temperature in the range of about 90°-100° C. to reduce said metal salt by said polysaccharide to colloidal metal(O) particles suspended in said polysaccharide solution and coated with said polysaccharide; (c) cooling the solution resulting from step (b) to room temperature and adding to the cooled solution an aqueous first bifunctional reagent capable of reacting with amine groups on said amine derivatized polysaccharide; (d) adding an organic diamine to the product of step (c); (e) reducing the reducible groups formed in step (d); (f) dialyzing the solution of step (e) against water; and (g) centrifuging the product of step (f) to obtain amine derivatized polysaccharide coated particles capable of being dispersed as colloids.
Claim 8
Independent Claim 179 words
A method of preparing colloidal metal(O) particles coated with an amine derivatized, oxidizable sugar containing polysaccharide, said method comprising: (a) heating an aqueous solution of a sugar reducible metal salt and adding to said solution an aqueous solution of an amine derivatized polysaccharide having at least one reducing sugar component; (b) heating and mixing the solution resulting from step (a) for about 10-30 minutes at a temperature in the range of about 90°-100° C. to reduce said metal salt by said polysaccharide to colloidal metal(O) particles suspended in said polysaccharide solution and coated with said polysaccharide; (c) cooling the solution resulting from step (b) to room temperature and adding to the cooled solution an aqueous first bifunctional reagent capable of reacting with amine groups on said amine derivatized polysaccharide; (d) adding an organic diamine to the product of step (c); (e) reducing the reducible groups formed in step (d); (f) dialyzing the solution of step (e) against water; and (g) centrifuging the product of step (f) to obtain amine derivatized polysaccharide coated particles capable of being dispersed as colloids.
Claim 9
Dependent Claim 12 words
The method of claim 6 wherein said derivatized polysaccharide is an aminodextran.
Claim 10
Dependent Claim 18 words
The method of claim 6 wherein said metal salt has a reduction potential of +0.7 volts or higher.
Claim 11
Dependent Claim 31 words
The method of claim 8 wherein said metal salt is selected from the group consisting of gold, silver, platinum, palladium, rhodium and iridium salts capable of being reduced by said polysaccharide.
Claim 12
Dependent Claim 20 words
The method of claim 9 wherein said metal salt is selected from the group consisting a gold or silver salt.
Claim 13
Dependent Claim 12 words
The method of claim 6 wherein said first bifunctional reagent is glutaraldehyde.
Claim 14
Dependent Claim 58 words
The method of claim 6 wherein said diamine is selected from the group consisting of H 2 NCH 2 --(CH 2 ) x --CH y (CH 3 ) z --NH 2 and C 6 H 4+a (NH 2 ) 2 , where x=0-3, y=1 or 2, and Z=1 when y=1 or Z=0 when y=2; and a=0 or 6.
Claim 15
Dependent Claim 60 words
A method of preparing colloidal metal(O) particles coated with an amine derivatized, oxidizable sugar containing polysaccharide and having a protein bound thereto, said method comprising: (a) preparing coated colloidal metal particles according to claim 1; and (b) reacting the particles of step (a) with a protein capable of forming a covalent bond with the amine groups present on said particles,
Claim 16
Dependent Claim 13 words
The method of to claim 13 wherein said protein is a monoclonal antibody.
Claim 17
Dependent Claim 28 words
The method according to claim 13 wherein said colloidal metal particles are selected from the group consisting of colloidal gold, silver, platinum, palladium, rhodium and iridium colloidal particles.
Claim 18
Dependent Claim 15 words
The method of claim 15 wherein said colloidal particles are colloidal gold or silver particles.
Claim 19
Dependent Claim 124 words
A method of preparing colloidal metal(O) particles coated with an amine-derivatized reducible sugar containing polysaccharide and having a protein bound thereto, said method comprising: (a) preparing colloidal metal particles according to claim 1; (b) converting the amine groups present on said particles to a group selected from the group consisting of sulhydryl and maleimidyl groups; and (c) reacting the product of step (b) with: (1) a protein having a functional group reactive with said sulfhydryl and maleimidyl groups or (2) a derivatized protein having at least one functional group selected from the group consisting of sulfhydryl and maleimidyl groups, such that when said particles contain sulfhydryl groups said proteins contains maleimidyl groups and when said particles contain maleimidyl groups said protein contains sulfhydryl groups.
Claim 20
Dependent Claim 49 words
The method of claim 17 wherein said protein is selected from the group consisting of a monoclonal and a polyclonal antibody either with or with a detectable label selected from the group consisting of an enzyme, a dye, an electron dense element and a substance containing a radioactive element.
Claim 21
Dependent Claim 27 words
The method of claim 17 wherein said colloidal metal particles are selected from the group consisting of colloidal gold, silver, platinum, palladium, rhodium and iridium colloidal particles.
Claim 22
Dependent Claim 15 words
The method of claim 19 wherein said colloidal particles are gold or silver colloidal particles.
Claim 23
Dependent Claim 53 words
Colloidal metal(O) particles having a polymeric organic coating thereon of a substance used to form said metal(O) particles by reduction of a metal salt with said substance, said coating comprising a stabilized amine derivatized polysaccharide and said coating being stabilized by means of a bifunctional crosslinking agent, a diamine and a reducing agent.
Claim 24
Dependent Claim 53 words
Colloidal metal(O) particles having a polymeric organic coating thereon of a substance used to form said metal(O) particles by reduction of a metal salt with said substance, said coating comprising a stabilized amine derivatized polysaccharide and said coating being stabilized by means of a bifunctional crosslinking agent, a diamine and a reducing agent.
Claim 25
Dependent Claim 13 words
The particles of claim 21 wherein said amine derivatized polysaccharide is an aminodextran.
Claim 26
Dependent Claim 12 words
The particles of claim 21 wherein said bifunctional crosslinking agent is glutaraldehyde.
Claim 27
Dependent Claim 58 words
The particles of claim 21 wherein said diamine is selected from the group consisting of H 2 NCH 2 --(CH 2 ) x --CH y (CH 3 ) z --NH 2 and C 6 H 4+a (NH 2 ) 2 , where x=0-3, y=1 or 2, and Z=1 when y=1 or Z=0 when y=2; and a=0 or 6.
Claim 28
Dependent Claim 22 words
The particles of claim 21 wherein said metal(O) is selected from the group consisting of gold, silver, platinum, palladium, rhodium and iridium.
Claim 29
Dependent Claim 12 words
The particle of claim 25 wherein said metal(O) is silver or gold.
Claim 30
Dependent Claim 63 words
Coated colloidal metal particles having a protein bound thereto, said particles comprising colloidal metal(O) particles coated with a stabilized amine derivatized polysaccharide and a protein bound to said coating, said coating being stabilized by means of a bifunctional crosslinking agent, a diamine and a reducing agent, and said metal(O) particles being formed by reduction of a metal salt with said amine derivatized polysaccharide.
Claim 31
Dependent Claim 63 words
Coated colloidal metal particles having a protein bound thereto, said particles comprising colloidal metal(O) particles coated with a stabilized amine derivatized polysaccharide and a protein bound to said coating, said coating being stabilized by means of a bifunctional crosslinking agent, a diamine and a reducing agent, and said metal(O) particles being formed by reduction of a metal salt with said amine derivatized polysaccharide.
Claim 32
Dependent Claim 13 words
The particles of claim 27 wherein said amine derivatized polysaccharide is an aminodextran.
Claim 33
Dependent Claim 12 words
The particles of claim 27 wherein said bifunctional crosslinking reagent is glutaraldehyde.
Claim 34
Dependent Claim 22 words
The particles of claim 27 wherein said metal(O) is selected from the group consisting of gold, silver, platinum, palladium, rhodium and iridium.
Claim 35
Dependent Claim 12 words
The particles of claim 30 wherein said metal(O) is gold or silver.
Claim 36
Dependent Claim 58 words
The particle of claim 27 wherein said diamine is selected from the group consisting of H 2 NCH 2 --(CH 2 ) x --CH y (CH 3 ) z --NH 2 and C 6 H 4+a (NH 2 ) 2 , where x=0-3, y=1 or 2, and Z=1 when y=1 or Z=0 when y=2; and a=0 or 6.
Claim 37
Dependent Claim 23 words
The particles according to claim 27 wherein said protein is selected from the group consisting of polyclonal antibodies, monoclonal antibodies, enzymes and lectins.
Claim 38
Dependent Claim 15 words
The particles according to claim 27 wherein said protein is a monoclonal or polyclonal antibody.
Claim 39
Dependent Claim 25 words
The particles according to claim 27 wherein said protein has a detectible enzyme or a substance containing a radioactive element or a dye attached thereto.
Claim 40
Dependent Claim 51 words
The particles according to claim 27 wherein said stabilized amine derivatized polysaccharide coated particles coated are reacted with selected reagents to produce particles having either reactive maleimidyl or sulfhydryl groups attached to said polysaccharide coating and said protein is covalently bound to the sulfhydryl or maleimidyl group on said coated particles.
Claim 41
Dependent Claim 57 words
Particles according to claim 36 wherein said protein is derivatized to contain either reactive sulfhydryl or maleimidyl groups prior to binding said protein to said maleimidyl or sulfhydryl containing polysaccharide coated particles, provided that when said particles contain sulfhydryl groups said protein contains maleimidyl groups and when said particles contain maleimidyl groups said protein contains sulfhydryl groups.

USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1996-09-27 1997-03-29 1997-09-28 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 2000-09-27 2001-03-29 2001-09-28 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 2004-09-27 2005-03-29 2005-09-28 $7700.00 Window Closed / Lapsed

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