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Ultrapurification of factor VIII using monoclonal antibodies

Assignee: Scripps Research Institute
•
Filing: 1983-12-21
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Granted: 1985-10-22
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Expires: 2003-12-21
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Abstract

No abstract record available.

Inventors

  • 👤 Theodore S. Zimmerman
  • 👤 Carol A. Fulcher

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-10-23 06:27
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/745; C07K14/755; A; A61; A61K; A61K38/00

Claims Analysis

Total Claims: 23
Claim 1
Independent Claim 70 words
An improved method of preparing Factor VIII procoagulant activity protein comprising the steps of (a) adsorbing a VIII:C/VIII:RP complex from a plasma or commercial concentrate source onto particles bound to a monoclonal antibody specific to VIII:RP, (b) eluting the VIII:C, (c) adsorbing the VIII:C obtained in step (b) in another adsorption to concentrate and further purify same, (d) eluting the adsorbed VIII:C, and (e) recovering highly purified and concentrated VIII:C.
Claim 2
Dependent Claim 70 words
An improved method of preparing Factor VIII procoagulant activity protein comprising the steps of (a) adsorbing a VIII:C/VIII:RP complex from a plasma or commercial concentrate source onto particles bound to a monoclonal antibody specific to VIII:RP, (b) eluting the VIII:C, (c) adsorbing the VIII:C obtained in step (b) in another adsorption to concentrate and further purify same, (d) eluting the adsorbed VIII:C, and (e) recovering highly purified and concentrated VIII:C.
Claim 3
Dependent Claim 21 words
A method according to claim 1, wherein the elutant used in each of steps (b) and (d) is a saline solution.
Claim 4
Dependent Claim 13 words
The method according to claim 2, wherein the saline solution is calcium chloride.
Claim 5
Dependent Claim 29 words
The method according to claim 3, wherein the concentration of said calcium chloride solution used in steps (b) and (d) ranges from about 0.25 M to about 0.5 M.
Claim 6
Dependent Claim 15 words
The method according to claim 1, wherein said adsorbent particles in step (a) are agarose.
Claim 7
Dependent Claim 17 words
The method according to claim 1, wherein aminohexyl agarose is employed in step (c) as the adsorbent.
Claim 8
Dependent Claim 50 words
The method according to claim 6, wherein calcium chloride solution is employed as the elutant in steps (b) and (d), concentration of said calcium chloride solution ranging from about 0.25 M to about 0.5 M in step (b) and from about 0.25 M to about 0.5 M in step (d).
Claim 9
Dependent Claim 22 words
An improved immunoadsorbent for isolation and purification of VIII:C from VIII:C/VIII:RP comprising a monoclonal antibody specific to VIII:RP bound to solid particles.
Claim 10
Dependent Claim 22 words
An improved immunoadsorbent for isolation and purification of VIII:C from VIII:C/VIII:RP comprising a monoclonal antibody specific to VIII:RP bound to solid particles.
Claim 11
Dependent Claim 13 words
The improved immunoadsorbent of claim 8, wherein said solid particles comprise a resin.
Claim 12
Dependent Claim 11 words
The improved immunoadsorbent of claim 9, wherein said resin comprises agarose.
Claim 13
Dependent Claim 12 words
The improved immunoadsorbent of claim 10, wherein said agarose is cross-linked agarose.
Claim 14
Dependent Claim 25 words
The improved immunoadsorbent of claim 11, wherein said immunoadsorbent has a coupling density of 3 to 4 g of monoclonal antibody per liter of agarose.
Claim 15
Dependent Claim 17 words
Highly purified and concentrated .Iadd.human or porcine .Iaddend.VIII:C prepared in accordance with the method of claim 1.
Claim 16
Dependent Claim 17 words
Highly purified and concentrated .Iadd.human or porcine .Iaddend.VIII:C prepared in accordance with the method of claim 6.
Claim 17
Dependent Claim 48 words
In a method for purifying Factor VIII procoagulant activity protein from plasma or concentrate, the improvement comprising the step of passing said plasma or concentrate through a chromatographic type column having adsorbent to which is bound monoclonal antibodies which is specific to VIII:RP and eluting the VIII-C therefrom.
Claim 18
Dependent Claim 48 words
In a method for purifying Factor VIII procoagulant activity protein from plasma or concentrate, the improvement comprising the step of passing said plasma or concentrate through a chromatographic type column having adsorbent to which is bound monoclonal antibodies which is specific to VIII:RP and eluting the VIII-C therefrom.
Claim 19
Dependent Claim 19 words
The method according to claim 15, wherein said adsorbent is agarose and said elutant is a saline solution. .Iadd.
Claim 20
Dependent Claim 133 words
A VIII:C of claim 13, wherein said VIII:C is human VIII:C. .Iaddend. .Iadd.18. A VIII:C of claim 14, wherein said VIII:C is human VIII:C. .Iaddend. .Iadd.19. An improved immunoadsorbent of claim 8 wherein said monoclonal antibody remains bound to VIII:RP during elution of VIII:C. .Iaddend. .Iadd.20. An improved immunoadsorbent of claim 19, wherein said elution is carried out with a saline solution. .Iaddend. .Iadd.21. An improved immunoadsorbent of claim 20 wherein said solution contains calcium ions. .Iaddend. .Iadd.22. An improved immunoadsorbent of claim 21, wherein said solution is calcium chloride. .Iaddend. .Iadd.23. An improved immunoadsorbent of claim 22, wherein the concentration of said calcium chloride is from 0.25 M to about 0.5 M. .Iaddend. .Iadd.24. A human VIII:C preparation having a potency in the range of 134 to 1172 units per ml, and
Claim 21
Dependent Claim 78 words
being substantially free of VIII:RP. .Iaddend. .Iadd.25. A human VIII:C preparation of claim 24, wherein the VIII:C concentration is at least 160,000 fold purified relative to VIII:C in plasma. .Iaddend. .Iadd.26. A human VIII:C preparation of claim 24, wherein the ratio of VIII:C to VIII:RP is greater than 100,000 times the ratio in plasma. .Iaddend. .Iadd.27. A human VIII:C preparation of claim 24, wherein said VIII:C is isolated from VIII:C/VIII:RP and 90-100 percent of the VIII:RP has been
Claim 22
Dependent Claim 112 words
removed. .Iaddend. .Iadd.28. A human VIII:C preparation having a specific activity greater than 2240 units/mg. .Iaddend. .Iadd.29. A human VIII:C preparation of claim 28 wherein the potency is in the range of 134 to 1172 units/ml. .Iaddend. .Iadd.30. An improved method of preparing Factor VIII procoagulant activity protein comprising the steps of (a) adsorbing a VIII:C/VIII:RP complex from a plasma or commercial concentrate source onto a substrate to which is bound a monoclonal antibody specific to VIII:RP, (b) eluting the VIII:C, (c) adsorbing the VIII:C obtained in step (b) in another adsorption to concentrate and further purify same, (d) eluting the adsorbed VIII:C, and (e) recovering highly purified and concentrated VIII:C. .Iaddend.
Claim 23
Dependent Claim 138 words
.Iadd. A method according to claim 30, wherein said substrate is a resin. .Iaddend. .Iadd.32. A method according to claim 30, wherein said substrate is comprised of particles. .Iaddend. .Iadd.33. An improved immunoadsorbent for isolation and purification of VIII:C from VIII:C/VIII:RP comprising a monoclonal antibody specific to VIII:RP bound to a substrate. .Iaddend. .Iadd.34. Highly purified and concentrated human or porcine VIII:C prepared in accordance with the method of claim 30. .Iaddend. .Iadd.35. In a method for purifying Factor VIII procoagulant activity protein from plasma or concentrate, the improvement comprising the step of passing said plasma or concentrate over a substrate to which is bound monoclonal antibodies which are specific to VIII:RP and eluting the VIII:C therefrom. .Iaddend. .Iadd.36. The method according to claim 35, wherein said substrate is agarose and said elutant is a saline solution. .Iaddend.

USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1988-10-21 1989-04-22 1989-10-22 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 1992-10-21 1993-04-22 1993-10-22 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 1996-10-21 1997-04-22 1997-10-22 $7700.00 Window Closed / Lapsed

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