43 results about "Recombinant factor viii" patented technology

Recombinant Factor VIII can be produced in relatively large quantities on a continuous basis from mammalian cells in the absence of any animal-derived proteins such as albumin by culturing the cells in a protein free medium supplemented with polyol copolymers, preferably in the presence of trace metals such as copper. In very preferred embodiments, the medium includes a polyglycol known as Pluronic F-68, copper sulfate, ferrous sulfate/EDTA complex, and salts of trace metals such as manganese, molybdenum, silicon, lithium and chromium. With an alternative medium which included trace copper ions alone (without polyol copolymers) we were also able to enhance the productivity of Factor VIII in recombinant cells such as BHK cells that are genetically engineered to express Factor VIII.

A method for purifying recombinant Factor VII (rFVII) or recombinant activated Factor VII (rFVIIa), comprising subjecting the rFVII or rFVIIa to liquid chromatography on a hydroxyapatite (HAP) column.

The invention relates to a recombinant factor VIII that includes one or more mutations at an interface of A1 and C2 domains of recombinant factor VIII. The one or more mutations include substitution of one or more amino acid residues with either a cysteine or an amino acid residue having a higher hydrophobicity. This results in enhanced stability of factor VIII. Methods for making the recombinant factor VIII, pharmaceutical compositions containing the recombinant factor VIII, and use of the recombinant factor VIII for treating hemophilia A are also disclosed.

The present invention relates to a recombinant factor VIII that is characterized by one or more mutations within a region surrounding an activated protein C cleavage site, which one or more mutations result in a reduced rate of inactivation by activated protein C. Isolated nucleic acid molecules, recombinant expression vectors, and host cells suitable for expression of the recombinant factor VIII are also disclosed. The recombinant factor VIII can be used for the treatment of clotting disorders, such as hemophilia A.

The present invention relates to recombinant factor VIII having a specific activity that is higher than that of the corresponding wild-type factor VIII. The present invention also relates to methods of making and using the recombinant factor VIII. The present invention also relates to an isolated nucleic acid molecule that encodes the recombinant factor VIII, as well as DNA expression systems and host cells containing the isolated nucleic acid molecule.

The invention provides a method for industrial-scale production of FVII polypeptides in mammalian cell culture free of animal-derived components.

Peptides that have domains that bind to recombinant factor VIII (rFVIII) are disclosed. A method of rFVIII binding assay using the peptides deduced from a combinatorial library in a filtration plate process is described. A method of using peptides having these available binding domains in an affinity chromatography process to purify factor VIII is also taught.

A method for purifying a recombinant protein using a multimodal or mixed mode resin containing ligands which comprise a hydrophobic part and a negatively charged part is described. The invention is advantageous in that it is a single step chromatographic process which does not require adjustment of pH or conductivity during loading step and results in high yield and potency. The process is used for the purification of recombinant compositions of coagulation factor, particularly recombinant Factor VIII.

The present invention relates to a recombinant Factor VIII molecule, wherein said molecule has reduced vWF binding capacity, and wherein said molecule is covalently conjugated with at least one side group.

The object of the present invention is to provide methods for the production of recombinant human Factor VIII, employing specific endoproteases, thus assuring full proteolytic processing of said factor even during its biosynthesis, consequently avoiding additional purification steps. Other objects of the present invention are the recombinant human Factor VIII as obtained by said methods, pharmaceutical compositions, related uses and therapeutic methods.

A recombinant human factor VIII or IX protein having a human glycosylation pattern but the protein is devoid of N-glycolylneuraminic acid and/or the carbohydrate group Galα-3Gal.

In one aspect, present invention provides a recombinant mutant human factor VIII having increased expression and/or secretion as compared to wild-type factor VIII. In certain embodiments, the recombinant factor VIII includes one or more amino acid substitution(s) selected from the group consisting of I86, Y105, A108, D115, Q117, F129, G132, H134, M147 and L152. In other aspects, the present invention provides FVIII encoding nucleic acids, FVIII-expression vectors, as well as methods of using the modified FVIII genes in the treatment of FVIII deficiencies, such as hemophilia A.

The present invention relates generally to a method of purifying Factor VII and/or Factor VIIa from a solution containing either or both proteins, the method comprising: diluting the solution containing Factor VII and/or Factor VIIa in a loading buffer; adding the diluted solution from step (i) to a multi-modal anion exchange resinunder conditions selected such that Factor VII and/or Factor VIIa is bound to the resin; optionally washing the resin with a wash buffer; adding an elution buffer to the resin under conditions selected such that Factor VII and/or Factor VIIa is eluted from the resin; and recovering the eluted recombinant Factor VII and/or Factor VIIa; wherein the loading buffer, the optional wash buffer and the elution buffer each comprise about 35 mM or less of calcium ions.

The present invention relates to the field of therapeutic proteins, in particular, to recombinant coagulation factors. It provides a recombinant Factor VIII (FVIII) protein comprising specific point mutations at defined positions, which serve to reduce the immunogenicity of said FVIII protein, wherein the Factor VIII protein substantially retains its coagulant activity. The invention further provides nucleic acids encoding said de-immunized protein, cell lines and methods of recombinant preparation as well as pharmaceutical compositions comprising the recombinant FVIII of the invention, whichare advantageous for use in treatment of patients with Hemophilia A, particularly those who have not yet been treated with a FVIII product. Additionally, it can be a safe alternative for previously treated patients and even for patients who have developed an immune-response to FVIII, e.g., for immune-tolerance-induction therapy (ITI/ITT) or rescue ITI. The invention also provides an assay for determining immunogenicity of a protein.

The invention relates to the field of bioengineering, in particular to a method for recombining and expressing a gene of a factor C from Tachypleus tridentatus in insect cells so as to detect and remove endotoxin. The method comprises the steps of preparing a recombinant rhabdovirus of the gene of the factor C from Tachypleus tridentatus, transfecting the insect cells, detecting the expression of the recombinant factor C from Tachypleus tridentatus in the insect cells, and the like. The recombinant factor C from Tachypleus tridentatus, prepared by the method, provides a precondition for researching the binding activity of endotoxin of the factor C, and also lays a foundation for the application of the factor C in detection and removal of the endotoxin.

The present invention relates to a method for treating gastrointestinal bleeding in a subject with severe von Willebrand Disease comprising administering to the subject at least one dose of recombinant von Willebrand Factor (rVWF) ranging from about 40 IU/kg to about 100 IU/kg, wherein the first dose further comprises recombinant Factor VIII (rFVIII).