Abstract: An improved method and device for measuring the mechanical properties of fibrin clots. A mixture of fibrinogen and thrombin is formed between two synthetic polymeric substrate materials and allowed to form a fibrin clot. The force required to separate the substrate materials is then measured and indicates the strength of the clot.

Abstract: An improved process for the purification of antibodies from human plasma or other sources is disclosed. The process involves suspension of the antibodies at pH 3.8 to 4.5 followed by addition of caprylic acid and a pH shift to pH 5.0 to 5.2. A precipitate of contaminating proteins, lipids and caprylate forms and is removed, while the majority of the antibodies remain in solution. Sodium caprylate is again added to a final concentration of not less than about 15 mM. This solution is incubated for 1 hour at 25.degree. C. to effect viral inactivation. A precipitate (mainly caprylate) is removed and the clear solution is diluted with purified water to reduce ionic strength. Anion exchange chromatography using two different resins is utilized to obtain an exceptionally pure IgG with subclass distribution similar to the starting distribution. The method maximizes yield and produces a gamma globulin with greater than 99% purity.

Abstract: An expression vector comprising a promoter, a coding sequence of a heterologous protein, the coding sequence being operably linked to the promoter, and an intronic sequence downstream of the promoter and upstream of the coding sequence, the intronic sequence comprising two identical donor sites and one acceptor site.

Abstract: A method and device for activating and applying a solution of fibrinogen to a desired site. The method includes contacting the solution of fibrinogen with immobilized thrombin resulting in an activated solution of polymerizable fibrin, and delivering the activated solution to the desired site. The device includes a housing having a compartment for a solution of fibrinogen, immobilized thrombin, a structure for bringing the solution of fibrinogen in contact with the immobilized thrombin under conditions which permit the activation of the fibrinogen resulting in polymerizable fibrin, and a structure for delivery of the activated solution to the desired site.

Abstract: A method of rendering a biologically active protein composition substantially free of viruses is disclosed. The method includes the steps of preparing lyophilized samples of the protein composition having a given moisture content, determining a minimum or threshold moisture content which, when the composition is heated to pasteurizing conditions will result in at least a 3.2 log.sub.10 reduction in virus titer, and then heating the sample under pasteurizing conditions that will result in at least a 3.2 log.sub.10 reduction in virus titer while maintaining biological activity of the protein.

Abstract: Peptide ligands which bind to thrombin and prothrombin are disclosed. The sequences of these peptides are Gln-Leu-Trp-Gly-Ser-His, Arg-Gln-Leu-Trp-Gly-Ser-His, His-Gln-Leu-Trp-Gly-Ser-His, and Tyr-Phe-Pro-Gly-Pro-Tyr-Leu. The preferred peptides have the sequence Gln-Leu-Trp-Gly-Ser-His or Tyr-Phe-Pro-Gly-Pro-Tyr-Leu. A method of using these peptides in an affinity chromatography process to purify thrombin is described.

Abstract: 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.

Abstract: A process for viral inactivation of a solution containing a biologically active protein, wherein the process comprises the steps of 1) contacting the solution with an immobilized ligand under conditions which allow protein to bind to the ligand, 2) subjecting the bound protein to a viral inactivation method under conditions which would result in substantial denaturation of the protein if it were not bound to the ligand, and 3) recovering the protein by washing the immobilized protein under conditions which favor the release of the protein into the solution under conditions in which the recovered protein retains its biological activity.

Abstract: Peptides which bind to fibrinogen are disclosed. These peptides have available fibrinogen binding domains having a Trp-Gln-Glu-His-Tyr-Asn, Trp-Gln-Glu-Thr-TyrGln,or Tyr-Glu-Asn-Tyr-Gly-Tyr sequence. Peptides containing at least one of these fibrinogen binding domains are immobilized upon a chromatographic substrate in a preferred embodiment of the invention. This preferred embodiment is useful in an affinity chromatography process to purify fibrinogen.

Abstract: Peptides which bind to fibrinogen are disclosed. These peptides have available fibrinogen binding domains having a Phe-Leu-Leu-Val or Leu-Leu-Val-Pro sequence. The preferred peptide is a 6-mer having the sequence Phe-Leu-Leu-Val-Pro-Leu (SEQ ID NO:1). A method of using these peptides in an affinity chromatography process to purify fibrinogen is described.

Abstract: Peptide ligands which bind to von Willebrand Factor (vWF) are disclosed. These peptides are selected from the group consisting of RLHSFY, RLKSFY, RLNSFY, RLQSFY, RFRSFY, RIRSFY, RVRSFY, RYRSFY, RLRSFY, HLRSFY, and KLRSFY. The preferred peptide ligand has the sequence RVRSFY. A method of using the disclosed peptides to purify vWF comprises binding the peptides to a support, then passing a solution containing vWF over the supports under condition such that the vWF will selectively bind to the peptides, and then eluting the vWF.

Abstract: Recombinant Factor VIII expression in a mammalian cell culture can be increased by including a novel liposome-like substance in the culture medium. The liposome-like substance comprises at least two (preferably at least three) different lipids in defined molar ratios. In a preferred embodiment, the addition of a liposome-like substance comprised of dioleoyl phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine in a molar ratio of 4:1:1 to the culture medium of GS-MDR cells resulted in an increase in FVIII production by a factor greater than five.

Abstract: A composition is disclosed which comprises a solution of human serum albumin essentially free of chemicals used in processing. The preparation is also essentially free of metals such as aluminum. The composition is 100% pure by cellulose acetate electrophoresis and is essentially monomeric when tested by high pressure liquid chromatography. The turbidity is less than 5 N.T.U. (National Turbidity Units). This preparation has a substantially longer shelf life and remains biologically active longer than products currently available. The novelty of this product is also such that it does not leach metallic substances such as aluminum from its closure. Novel applications of process methodology are taught in the preparation of this composition and a novel preparation results from essentially non hemoglobin containing albumin sources such as Source Plasma (Human).