The invention relates to commercially viable methods for producing biologically active vitamin R dependent proteins, particularly Factor IX. Factor IX is produced at a level of at least about 15 mg/L and is at least 25% biologically active. The method relies upon co-expression of one or more of paired basic amino acid converting enzyme (PACE), vitamin K dependent epoxide reductase (VKOR) and vitamin K dependent -glutamyl carboxylase (VKGC) at a preferred ratio so that the vitamin K dependent protein is efficiently produced and processed by a recombinant cell.

The present invention is directed to compounds useful in stabilizing thrombin activity, thrombin compositions comprising the compounds, methods of using the compounds and methods of identifying compounds capable of stabilizing thrombin activity. The compounds are preferably isolated peptides comprising or interacting with the gamma loop of thrombin.

The present invention is directed to compounds, methods for stabilizing thrombin activity with a thrombin binding oligonucleotide and to stabilized thrombin. The thrombin binding oligonucleotide is capable of inhibiting thrombin activity whereby the inhibition can be reversed with an antisense oligonucleotide.

A system for preparing a thrombin serum that can include a containment device, a cage received within the containment device, a cap attachable to the containment device, an inlet port configured to introduce a non-anti-coagulated autologous blood fluid into the containment device, and an outlet port. An activator, such as glass beads, can be present within the containment device.

A device, interface complex, diagnostic system, kit or method for use in binding analyte of interest, wherein immobilizing is on an aluminum oxide surface. An interface molecule is immobilized on the aluminum oxide surface. Attached to the interface molecule, is a cross linking agent for binding to the analyte, or a biomolecule specific to the analyte. The interface molecule includes a polypeptide having at least one carboxy rich domain providing at least 5 free carboxyl groups within a molecular volume of 2.2-25 nm.sup.3, the free carboxyl groups being provided by amino acids containing two or more carboxyl groups, through which the interface molecule is immobilized to the aluminum oxide surface. The biomolecule may be covalently attached to the interface molecule, or the biomolecule may bean engineered antibody attached to the interface molecule through an antigenic determinant or through an Fc fragment.

The present invention relates to recombinant proteins having serine protease polypeptides that have serine protease activity in the presence of a serine protease inhibitor and that are able to completely or partially reverse a serine protease inhibitor effect, for example in a subject treated with a serine protease inhibitor. More specifically, described herein are recombinant proteins and methods for completely or partially reversing an anti-coagulant effect of a coagulation inhibitor.

Aspects of the disclosure relate to methods and compositions useful for in vivo and/or in vitro enzyme profiling. In some embodiments, the disclosure provides methods of in vivo enzymatic processing of exogenous molecules followed by detection of signature molecules as representative of the presence of active enzymes associated with diseases or conditions. In some embodiments, the disclosure provides compositions and in vitro methods for localization of enzymatic activity in a tissue sample.

The present invention is directed to compounds useful in stabilizing thrombin activity, thrombin compositions comprising the compounds, methods of using the compounds and methods of identifying compounds capable of stabilizing thrombin activity. The compounds are preferably isolated peptides comprising or interacting with the gamma loop of thrombin.

A method of determining generation of an activated protease in a biological sample is provided. The method comprises the steps of exposing a biological sample to a substrate for the activated protease, wherein the substrate comprises a detectable label linked to a cleavage sequence for the activated protease by C-terminal and N-terminal spacers that form a beta-sheet, and wherein the detectable label emits a first signal associated with the substrate and second signal associated with a cleaved product; and determining the generation of the activated protease by measuring the change in the first or second signal over time.

During the process of coagulation, prothrombin is activated to ?-thrombin by prothrombinase. Key residues in the structure of prothrombin allow for modulation of the activation of prothrombin. In certain embodiments, a recombinant prothrombin with at least one point mutation or deletion is provided.