41 results about "Blood coagulation factor IX" patented technology

The present inventors succeeded in constructing bispecific antibodies, which bind to both the blood coagulation factor IX/activated blood coagulation factor IX and blood coagulation factor X, and functionally substitute for blood coagulation factor VIII/activated blood coagulation factor VIII which enhances the enzymatic reaction.

Various bispecific antibodies that specifically bind to both blood coagulation factor IX/activated blood coagulation factor IX and blood coagulation factor X and functionally substitute for the cofactor function of blood coagulation factor VIII, that is, the function to promote activation of blood coagulation factor X by activated blood coagulation factor IX, were produced. From these antibodies, multispecific antigen-binding molecules having a high activity of functionally substituting for blood coagulation factor VIII were successfully discovered.

The present invention relates to recombinant blood coagulation factor IX (rFIX) mutants having improved FIX clotting activity. Three full length FIX proteins with combinations of mutations of amino acids important for functional activity of FIX and FIX wild type were cloned and expressed in HEK 293 cells. The proteins were tested by an activated partial thromboplastin time (aPTT) assays in FIX-depleted plasma. Two mutant proteins had increased specific FIX activity. Furthermore, a pre-activated FIX protein had an increased activity in FIX-depleted plasma. Therefore these FIX mutants can be used for the treatment of FIX associated bleeding disorders.

The invention discloses a monoclonal antibody of a blood coagulation factor IX. The antibody comprises a light chain and a heavy chain, wherein the variable region amino acid sequence of the light chain is represented by SEQ ID NO.3, and the variable region amino acid sequence of the heavy chain is represented by SEQ ID NO.4. The invention also discloses a preparation method of the antibody. The antibody can specifically identify the blood coagulation factor IX. The invention further discloses a method for purifying the blood coagulation factor IX from blood plasma through utilizing the antibody. The blood coagulation factor IX can be conveniently and efficiently purified from the blood plasma through utilizing the antibody, so the infection risks of the diseases comprising hepatitis A, the hepatitis B, AIDS and the like caused by the patient blood transfusion supplementation of the blood coagulation factor IX are reduced.

The invention relates to preparation and application of high-activity blood coagulation factor IX mutant, recombinant protein and fusion protein. The nucleotide sequence of the mutant is as shown in SEQ ID No. 1.

The invention relates to a preparation method of a clinical blood coagulation inspection preparation and particularly relates to a preparation method of a blood coagulation factor IX quality control product. The preparation method comprises the following steps: carrying out affinity chromatography on the mixed blood plasma of multiple persons by using an anti-human blood coagulation factor IX monoclonal antibody immunoaffinity chromatography column, removing a blood coagulation factor IX in the mixed blood plasma of multiple persons to obtain a blood plasma in shortage of the blood coagulation factor IX; mixing the mixed blood plasma of multiple persons with the blood plasma in shortage of the blood coagulation factor IX according to a certain proportion to prepare a blood coagulation factor IX quality control product with the content of the blood coagulation factor IX at different concentration levels, adding a freeze-drying protective additive, carrying out sub-packaging, and carrying out freeze drying so as to obtain the blood coagulation factor IX quality control product. According to the blood coagulation factor IX quality control product prepared by the preparation method, the uniformity, the stability and the stability of freeze-dried aquatic product subjected to re-melting are good, and the quality control product can replace an imported product to be used for quality control on detection of blood coagulation factor IX, so that the reduction of the detection cost is facilitated and the capability of detecting the blood coagulation factor IX in China can be promoted.

The invention relates to the field of medicinal biotechnology, and in particular to a high-purity prothrombin complex product stabilizer capable of effectively preventing the inactivation of a blood coagulation factor IX with a main effect and blood coagulation factors II, VII and X in a product in a freeze-drying process, wherein the specific activity of each of four blood coagulation factors II, VII, IX and X is greater than or equal to 3.5IU/mg protein. The stabilizer provided by the invention contains arginine or/and trehalose or/and histidine or/and glycine. The tests show that so long as the high-purity prothrombin complex contains 0.1-10% of arginine or/and 0.1-10% of trehalose or/and 0.1-10% of histidine, the activity of the blood coagulation factor IX and the activity of the blood coagulation factors II, VII and X can be effectively protected in the freeze-drying process. Thus, the stabilizer provided by the invention can be applied to the freeze-drying process of the high-purity prothrombin complex.

The invention provides a method for efficiently extracting and purifying a blood coagulation factor IX and a blood coagulation factor X. The method provided by the invention comprises the following steps: collecting blood, treating, and collecting blood plasma; and extracting an IX crude extract and an X crude extract from the blood plasma, and purifying. The method is characterized in that the purifying step is to use FIX/FX-bp-Sepharose 4B to purify the blood coagulation factor IX crude extract and the blood coagulation factor X crude extract respectively through the affinity chromatography; and the affinity ligand FIX/FX-bp is selected from ACF I, ACF II or AHP. The purified blood coagulation factor IX can be directly used as a medicament and the purified blood coagulation factor X canbe directly used as a reagent.

The invention relates to a genetically-modified non-human animal and application thereof, in particular to a transgenic mouse model and application thereof in the aspect of screening of a targeted medicine. According to the transgenic mouse model and application thereof in the aspect of screening of the targeted medicine, by targetedly knocking out and replacing a mouse protein C gene by means ofa human protein C gene expression cassette, a humanized protein C knock-in mouse is generated. The mouse has fertility, and can hybridize with another mouse disease model (such as a mouse in the deficiency of a blood coagulation factor VIII or a blood coagulation factor IX) to produce a humanized protein C mouse disease model (such as a humanized protein C factor VIII-deficient mouse model or a humanized protein C factor IX-deficient mouse model). The mouse model can be used for studying functions in vivo of human protein C and human activated protein C (APC). The mouse model is the first mouse model used for testing a therapeutic candidate medicine, which targets the human protein C or the APC, in vivo, and has very high economic value and scientific research value.

The invention discloses a method for detecting anticoagulant capacity of a human prothrombin complex. The method comprises the following steps: (1) taking the human prothrombin complex, diluting until a blood coagulation factor IX is 0.8-1.3IU/ml, adding a human prothrombin solution with the isovolumetric concentration being 4-6IU/ml, mixing evenly, and standing at room temperature for 1-2 minutes; (2) adding a thrombin chromogenic substrate solution which is isovolumetric to the human prothrombin solution, mixing evenly, and incubating at 22-28 DEG C for 4-6 minutes, wherein the concentration of the chromogenic substrate solution is 0.5-0.7mg/ml; (3) determining a light absorption value at the wavelength of 405nm, and determining once 30-50 seconds for 5-8 times in all; and (4) drawing the time changing trend along with the light absorption value by taking the determined light absorption value as a longitudinal coordinate (y) and time (s) as a cross coordinate (x), building a linear equation y=kx+a, wherein k is slope, and calculating the value of 1/k. The method for detecting the anticoagulant capacity of the human prothrombin complex is simple and convenient to operate and good in repeatability; the comprehensive anticoagulant ability of the PCCs product can be quantitatively detected; and the safety of the PCCs product can be evaluated.

The inventors discovered that by administering a pharmaceutical composition comprising a bispecific antigen-binding molecule that recognizes blood coagulation factor IX and/or activated blood coagulation factor IX and blood coagulation factor X and/or activated blood coagulation factor X according to a given dosage regimen, diseases that develop and/or progress due to a decrease or deficiency in the activity of blood coagulation factor VIII and/or activated blood coagulation factor VIII can be prevented and/or treated more effectively.

The present invention relates to a blood coagulation factor IX fusion protein having an extended circulating half-life, a conjugate containing the fusion protein, a pharmaceutical composition containing the fusion protein and the conjugate and an application of the fusion protein, the conjugate and the composition in the treatment of hemorrhagic diseases (such as hemophilia B).