US5200510A
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Method for purifying factor viii:c, von willebrand factor and complexes thereof
Assignee: Zymogenetics Inc
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Filing: 1988-03-02
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Granted: 1993-04-06
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Expires: 2008-03-02
Abstract
Abstract Methods for purifying factor VIII:C, von Willebrand factor (vWF) or complexes thereof from heterogeneous biological fluids are disclosed. The methods utilize a binding peptide, specific to either factor VIII:C or vWF, bound to an insoluble matrix. Peptides suitable for use within the methods are also disclosed.
Inventors
- 👤 Anur A. Kumar
- 👤 Frederick S. Hagen
- 👤 Andrzej Z. Sledziewski
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 17:10
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/745; C07K14/755
Claims Analysis
Total Claims: 25
Claim 1
Independent Claim
91 words
A method for purifying von Willebrand factor from a heterogeneous biological fluid, comprising: exposing the biological fluid to a peptide that specifically binds to von Willebrand factor, said peptide consisting essentially of between four and ninety-six amino acids and containing a sequence of at least four consecutive amino acids corresponding to amino acids 165-260 of glycoprotein Ib, said peptide bound to an insoluble matrix such that the von Willebrand factor specifically binds to said peptide; eluting the bound von Willebrand factor from the peptide; and collecting the von Willebrand factor-containing eluate.
Claim 2
Independent Claim
91 words
A method for purifying von Willebrand factor from a heterogeneous biological fluid, comprising: exposing the biological fluid to a peptide that specifically binds to von Willebrand factor, said peptide consisting essentially of between four and ninety-six amino acids and containing a sequence of at least four consecutive amino acids corresponding to amino acids 165-260 of glycoprotein Ib, said peptide bound to an insoluble matrix such that the von Willebrand factor specifically binds to said peptide; eluting the bound von Willebrand factor from the peptide; and collecting the von Willebrand factor-containing eluate.
Claim 3
Dependent Claim
19 words
The method of claim 1 including, subsequent to the step of exposing, washing non-specifically bound elements from the matrix.
Claim 4
Dependent Claim
17 words
The method of claim 1 including, subsequent to the step of collecting, concentrating the von Willebrand factor.
Claim 5
Dependent Claim
36 words
The method of claim 1 wherein the step of eluting comprises exposing the bound von Willebrand factor to a pH gradient or a buffer having a salt concentration sufficient to elute said bound von Willebrand factor.
Claim 6
Dependent Claim
36 words
The method of claim 1 wherein said peptide consists of between approximately four and forty amino acids and comprises a sequence of at least four consecutive amino acids corresponding to amino acids 165-260 of glycoprotein Ib.
Claim 7
Dependent Claim
22 words
The method of claim 1 wherein said peptide is selected from the group consisting of PEP-12, PEP-13, PEP-14, PEP-15, PEP-16 and PEP-17.
Claim 8
Independent Claim
100 words
A method for purifying a factor VIII:C-von Willebrand factor complex from a heterogeneous biological fluid, comprising: exposing the biological fluid to a peptide that specifically binds to von Willebrand factor, said peptide consisting essentially of between four and ninety-six amino acids and containing a sequence of at least four consecutive amino acids corresponding to amino acids 165-260 of glycoprotein Ib, said peptide bound to an insoluble matrix such that the factor VIII:C-von Willebrand factor complex specifically binds to said peptide; eluting the bound factor VIII:C-von Willebrand factor complex from the peptide; and collecting the factor VIII:C-von Willebrand factor complex-containing eluate.
Claim 9
Independent Claim
100 words
A method for purifying a factor VIII:C-von Willebrand factor complex from a heterogeneous biological fluid, comprising: exposing the biological fluid to a peptide that specifically binds to von Willebrand factor, said peptide consisting essentially of between four and ninety-six amino acids and containing a sequence of at least four consecutive amino acids corresponding to amino acids 165-260 of glycoprotein Ib, said peptide bound to an insoluble matrix such that the factor VIII:C-von Willebrand factor complex specifically binds to said peptide; eluting the bound factor VIII:C-von Willebrand factor complex from the peptide; and collecting the factor VIII:C-von Willebrand factor complex-containing eluate.
Claim 10
Dependent Claim
19 words
The method of claim 7 including, subsequent to the step of exposing, washing non-specifically bound elements from the matrix.
Claim 11
Dependent Claim
19 words
The method of claim 7 including, subsequent to the step of collecting, concentrating the factor VIII:C-von Willebrand factor complex.
Claim 12
Dependent Claim
21 words
The method of claim 7 wherein the step of eluting comprises exposing the bound von Willebrand factor to a pH gradient.
Claim 13
Dependent Claim
36 words
The method of claim 7 wherein said peptide consists of between approximately four and forty amino acids and comprises a sequence of at least four consecutive amino acids corresponding to amino acids 165-260 of glycoprotein Ib.
Claim 14
Dependent Claim
22 words
The method of claim 7 wherein said peptide is selected from the group consisting of PEP-12, PEP-13, PEP-14, PEP-15, PEP-16 and PEP-17.
Claim 15
Independent Claim
95 words
A method for purifying factor VIII:C from a heterogeneous biological fluid containing factor VIII:C-von Willebrand factor complexes, comprising: exposing the biological fluid to a peptide that specifically binds to von Willebrand factor said peptide consisting essentially of between four and ninety-six amino acids and containing a sequence of at least four consecutive amino acids corresponding to amino acids 165-260 of glycoprotein Ib, said peptide bound to an insoluble matrix, such that the factor VIII:C-von Willebrand factor complex specifically binds to said peptide; eluting the factor VIII:C from the complex; and collecting the factor VIII:C-containing eluate.
Claim 16
Independent Claim
95 words
A method for purifying factor VIII:C from a heterogeneous biological fluid containing factor VIII:C-von Willebrand factor complexes, comprising: exposing the biological fluid to a peptide that specifically binds to von Willebrand factor said peptide consisting essentially of between four and ninety-six amino acids and containing a sequence of at least four consecutive amino acids corresponding to amino acids 165-260 of glycoprotein Ib, said peptide bound to an insoluble matrix, such that the factor VIII:C-von Willebrand factor complex specifically binds to said peptide; eluting the factor VIII:C from the complex; and collecting the factor VIII:C-containing eluate.
Claim 17
Dependent Claim
19 words
The method of claim 13 wherein said peptide consists of between approximately four amino acids and forty amino acids.
Claim 18
Dependent Claim
15 words
The method of claim 13 wherein said peptide includes a terminal lysine or cysteine residue.
Claim 19
Dependent Claim
19 words
The method of claim 13 including, subsequent to the step of exposing, washing non-specifically bound elements from the matrix.
Claim 20
Dependent Claim
16 words
The method of claim 13 including, subsequent to the step of collecting, concentrating the factor VIII:C.
Claim 21
Dependent Claim
27 words
The method of claim 13 wherein the step of eluting comprises exposing the bound complex to a solution having an ionic strength sufficient to separate said complex.
Claim 22
Dependent Claim
36 words
The method of claim 13 wherein said peptide consists of between approximately four and forty amino acids and comprises a sequence of at least four consecutive amino acids corresponding to amino acids 165-260 of glycoprotein Ib.
Claim 23
Dependent Claim
22 words
The method of claim 13 wherein said peptide is selected from the group consisting of PEP-12, PEP-13, PEP-14, PEP-15, PEP-16 and PEP-17.
Claim 24
Independent Claim
102 words
A method for purifying factor VIII:C from a heterogeneous biological fluid containing factor VIII:C-von Willebrand factor complexes, comprising: exposing the biological fluid to a peptide that specifically binds to von Willebrand factor, said peptide consisting essentially of between four and ninety-six amino acids and containing a sequence of at least four consecutive amino acids corresponding to amino acids 165-260 of glycoprotein Ib, said peptide being bound to an insoluble matrix, such that the factor VIII:C-von Willebrand factor complex specifically binds to the peptide; eluting the bound complexes from the peptide; dissociating the factor VIII:C-von Willebrand factor complex; and isolating the factor VIII:C.
Claim 25
Independent Claim
102 words
A method for purifying factor VIII:C from a heterogeneous biological fluid containing factor VIII:C-von Willebrand factor complexes, comprising: exposing the biological fluid to a peptide that specifically binds to von Willebrand factor, said peptide consisting essentially of between four and ninety-six amino acids and containing a sequence of at least four consecutive amino acids corresponding to amino acids 165-260 of glycoprotein Ib, said peptide being bound to an insoluble matrix, such that the factor VIII:C-von Willebrand factor complex specifically binds to the peptide; eluting the bound complexes from the peptide; dissociating the factor VIII:C-von Willebrand factor complex; and isolating the factor VIII:C.
References Cited (Backward Citations)
USRE32011E
Cited
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