US5204447A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Purification of factor xiii
Assignee: Zymogenetics Inc
•
Filing: 1988-11-14
•
Granted: 1993-04-20
•
Expires: 2008-11-14
Abstract
Abstract Methods for purifying factor XIII from a biological fluid are provided. The methods comprise precipitation of factor XIII by adjusting the pH of the biological fluid to 5.5 to 6.5 and recovering the precipitated factor XIII.
Inventors
- 👤 Paul D. Bishop
- 👤 Gerald W. Lasser
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-25 23:24
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/10; C12N9/1025; C12N9/104; C12N9/1044
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
33 words
A method for purifying factor XIII from a biological fluid, comprising: precipitating the factor XIII by adjusting the pH of the biological fluid to 5.5 to 6.5; and recovering the precipitated factor XIII.
Claim 2
Independent Claim
33 words
A method for purifying factor XIII from a biological fluid, comprising: precipitating the factor XIII by adjusting the pH of the biological fluid to 5.5 to 6.5; and recovering the precipitated factor XIII.
Claim 3
Dependent Claim
25 words
The method of claim 1 wherein the pH of the fluid is adjusted by the use of a low ionic strength, polyamine or phosphate buffer.
Claim 4
Dependent Claim
22 words
The method of claim 2 wherein the low ionic strength polyamine is selected from the group consisting of piperazine and derivatives thereof.
Claim 5
Dependent Claim
23 words
The method of claim 1 wherein the precipitating step comprises dialyzing the biological fluid in a buffer comprising 10-100 mM piperazine, pH 6.0.
Claim 6
Dependent Claim
13 words
The method of claim 1, further comprising washing the recovered, precipitated factor XIII.
Claim 7
Independent Claim
46 words
A method for purifying factor XIII from a biological fluid, comprising: fractioning the biological fluid by anion exchange chromatography to produce an enriched fraction; precipitating the factor XIII by adjusting the pH of the enriched fraction to 5.5 to 6.5; and recovering the precipitated factor XIII.
Claim 8
Independent Claim
46 words
A method for purifying factor XIII from a biological fluid, comprising: fractioning the biological fluid by anion exchange chromatography to produce an enriched fraction; precipitating the factor XIII by adjusting the pH of the enriched fraction to 5.5 to 6.5; and recovering the precipitated factor XIII.
Claim 9
Dependent Claim
37 words
The method of claim 6, comprising the additional steps of: dissolving the recovered, precipitated factor XIII to produce a solution; fractionating the solution by chromatography to produce a second enriched fraction; and recovering the second enriched fraction.
Claim 10
Dependent Claim
26 words
The method of claim 6 wherein the pH of the enriched fraction is adjusted by the use of a low ionic strength, polyamine or phosphate buffer.
Claim 11
Dependent Claim
22 words
The method of claim 8 wherein the low ionic strength polyamine is selected from the group consisting of piperazine and derivatives thereof.
Claim 12
Dependent Claim
23 words
The method of claim 6 wherein the precipitating step comprises dialyzing the enriched fraction in a buffer comprising 10-100 mM piperazine, pH 6.0.
Claim 13
Dependent Claim
14 words
The method of claim 6 wherein the biological fluid is a cleared cell lysate.
Claim 14
Dependent Claim
15 words
The method of claim 11 wherein the cleared cell lysate is a yeast cell lysate.
Claim 15
Dependent Claim
53 words
The method of claim 6 wherein the biological fluid is prepared from a cell lysate by: removing particulate material from a cell lysate to produce a cleared lysate; adding a precipitating agent to the cleared lysate to produce a precipitate; and resuspending the precipitate in a suitable buffer to form a biological fluid.
Claim 16
Dependent Claim
40 words
The method of claim 13 wherein the step of adding a precipitating agent comprises adding ammonium sulfate to between about 30% to 40% of saturation, or adding polyethylene glycol to a concentration of between about 6% to 17% by weight.
Claim 17
Dependent Claim
16 words
The method of claim 6 wherein the fractionating step comprises chromatography on DEAE anion exchange media.
Claim 18
Dependent Claim
24 words
The method of claim 7 wherein the step of fractionating the solution by chromatography comprises filtration of the solution on gel filtration chromatography media.
Claim 19
Independent Claim
106 words
A method for purifying factor XIII from a biological fluid, comprising: adding to a biological fluid ammonium sulfate to about 30% to about 40% of saturation to produce a precipitate; resuspending the precipitate in a low ionic strength, mildly alkaline buffer; fractioning the resuspended precipitate by anion exchange chromatography to produce an enriched fraction; precipitating the factor XIII by adjusting the pH of the enriched fraction to 5.5 to 6.5 with piperazine buffer; recovering the precipitated factor XIII; dissolving the precipitated factor XIII to produce a solution; fractioning the solution by size exclusion chromatography to produce a second enriched fraction; and recovering the second enriched fraction.
Claim 20
Independent Claim
106 words
A method for purifying factor XIII from a biological fluid, comprising: adding to a biological fluid ammonium sulfate to about 30% to about 40% of saturation to produce a precipitate; resuspending the precipitate in a low ionic strength, mildly alkaline buffer; fractioning the resuspended precipitate by anion exchange chromatography to produce an enriched fraction; precipitating the factor XIII by adjusting the pH of the enriched fraction to 5.5 to 6.5 with piperazine buffer; recovering the precipitated factor XIII; dissolving the precipitated factor XIII to produce a solution; fractioning the solution by size exclusion chromatography to produce a second enriched fraction; and recovering the second enriched fraction.
Claim 21
Dependent Claim
14 words
The method of claim 17 wherein the biological fluid is a cleared cell lysate.
Claim 22
Dependent Claim
15 words
The method of claim 18 wherein the cleared cell lysate is a yeast cell lysate.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1996-04-19 | 1996-10-19 | 1997-04-20 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2000-04-19 | 2000-10-19 | 2001-04-20 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2004-04-19 | 2004-10-19 | 2005-04-20 | $7700.00 | Window Closed / Lapsed |