35 results about "ADAMTS13" patented technology

Provided herein are methods for purifying recombinant A Disintegrin-like and Metallopeptidase with Thrombospondin Type 1 Motif 13 (ADAMTS13) protein from a sample. The method comprises enriching for ADAMTS13 protein by chromatographically contacting the sample with hydroxyapatite under conditions that allow ADAMTS13 protein to appear in the eluate or supernatant from the hydroxylapatite. The methods may further comprise tandem chromatography with a mixed mode cation exchange/hydrophobic interaction resin that binds ADAMTS13 protein. Additional optional steps involve ultrafiltration/diafiltration, anion exchange chromatography, cation exchange chromatography, and viral inactivation. Also provided herein are methods for inactivating virus contaminants in protein samples, where the protein is immobilized on a support. Also provided herein are compositions of ADAMTS13 prepared according to said methods.

The present invention provides culture mediums that are useful for the expression of ADAMTS proteins, such as ADAMTS13. Methods for the expression and purification of ADAMTS proteins are also provided. In some embodiments, the mediums and methods of the invention are useful for the expression of ADAMTS proteins having high specific activities. Also provided are ADAMTS, e.g., ADAMTS13, protein compositions with high specific activities, which are expressed and purified according to the methods provided herein.

This invention relates to methods for treating or preventing an infarction by administering to a patient in need thereof a compound capable of suppressing the expression or activity of the von Willebrand Factor (VWF). Thus, the invention relates to the use of a pharmaceutically effective amount of a VWF inhibitor, such as ADAMTS13, for the preparation of a medicament for treating conditions known to involve infarction to reduce or eliminate the symptoms and effect of an infarction.

Provided are assays and kits to detect ADAMTS13 activity using peptide substrates and ADAMTS13/Factor XI complexes using ELISA. These assays and kits can be used for diagnostic applications and to evaluate treatment of thrombotic or hemostatic disorders, for example, thrombotic thrombocytopenic purpura (TTP). Also provided is a novel form of ADAMTS13 found on platelets, and anti-ADAMTS13 antibodies.

Methods of analyzing the electrophoretic mobility distribution of von Willebrand factor (vWF) multimers include providing a sample medium comprising a plurality vWF multimers. The vWF multimers are electrophoretically separated by electrohoretic mobility in the sample medium by subjecting said sample medium to an electric field to provide separated vWF multimers. The separated vWF multimers in the sample medium are exposed to a light source to produce scattered light. The scattered light is detected, and the electrophoretic mobility distribution of the separated vWF multimers is determined from the detected scattered light.

The invention belongs to the technical field of biology, in particular to a fluorogenic substrate for detecting ADAMTS13 enzymatic activity and a detection method. The fluorogenic substrate for detecting the ADAMTS13 enzymatic activity comprises an amino acid sequence shown as SEQ ID NO:2, and sulfydryl of cysteine at the 4 position and the 15 position at the N end of the amino acid sequence is combined with fluorescein-5- maleimide. Q1599C of the fluorogenic substrate is closer to N1611C modified site, simultaneously the N-end sequence of the fluorogenic substrate does not contain extra amino acid GS, and enzymolysis can be conducted on the fluorogenic substrate through ADAMTS13 more effectively. Therefore, sensitivity for detecting ADAMTS13 enzymatic activity can be improved. The method for detecting the ADAMTS13 enzymatic activity is convenient to operate, accurate in detection result, high in sensitivity and suitable for quick detection of the ADAMTS13 enzymatic activity in blood plasma in hospitals, health departments and medical research and development institutions at different levels.

The invention relates to the field of use of a biomarker for a disease, and discloses biomarkers von willebrand Factor (VWF) and ADAMTS13 and use thereof in a cirrhosis diagnosis reagent. The VWF disclosed by the invention is formed by protein of which the accession number of an amino acid sequence in a database for the national center of biotechnology information (NCBI) is GI317373549; the ADAMTS13 is formed by protein of which the accession number in the database for the NCBI is GI74749836. A testee is subjected to cirrhosis diagnosis by the biomarkers VWF and ADAMTS13, and the biomarkers have the advantages of being simple and feasible, safe and free of a wound in the diagnosis process, easy to be accepted by a patient, small in effect on diagnostic criteria unification caused by personal subjective factors, high in specificity and sensitivity on diagnosis result, easy to carry out a lot of screening for selection, and the like.

The present invention relates to methods and means for detecting ADAMTS13 immune complexes in a sample. The methods include the steps of capturing and labelling immune complexes of anti-ADAMTS13 antibodies. Capturing and labelling may be achieved by two different binding units targeting the immune complexes. The invention further relates to diagnosing diseases associated with immunologic ADAMTS13 dysfunction like TTP (thrombotic thrombocytopenic purpura).

The present invention further relates to specific humanised monoclonal antibodies inhibiting ADAMTS13 function. The present invention relates to methods of treatment by human ADAMTS13 inhibition in circulatory assist device induced haemorrhagic complication such as a bleeding disorder, in particular, bleeding after left ventricular assist device (LVAD) implantation.

The present invention relates to methods of treatment by human ADAMTS13 inhibition in circulatory assist device induced haemorrhagic complication such as a bleeding disorder, in particular, bleeding after left ventricular assist device implantation. The present invention further relates to specific monoclonal antibodies inhibiting ADAMTS13 function.

The invention relates to an ADAMTS13 substrate with histidine tags. An amino acid sequence of the substrate is as shown in SEQ ID No.1 and the substrate also has activity of protein enzyme-digested byADAMTS13. The ADAMTS13 enzyme-digested substrate provided by the invention can be efficiently expressed and purified in vitro in a large quantity, does not need to be subjected to fluorescent modification to further reduce cost, and can be widely applied to actual clinic. Besides, the substrate respectively carrying two groups of different tags can be used as a tag tool for protein purification,besides, two products produced after enzyme digestion with the ADAMTS13 respectively have tags with different specificities, and the two products can both be detected by an ELISA kit for the tags. Thenovel ADAMTS13 enzyme-digested substrate provided by the invention can be efficiently enzyme-digested by the ADAMTS13 and can be applied to detection of activity of the ADAMTS13.

The invention discloses an inhibitory monoclonal antibody resistant to lyase of a human von Willebrand factor (VWF) and application of the inhibitory monoclonal antibody. The monoclonal antibody belongs to an IgG1 subclass antibody and is produced from a hybridoma cell strain SZ-169; the preservation number of the hybridoma cell strain SZ-169 is CCTCC NO: C2016134. Compared with the prior art, the inhibitory monoclonal antibody has the following advantages: (1) the monoclonal antibody can be specifically combined with human overall-length ADAMTS13 and truncated ADAMTS13-T7 proteins, and the bonding capacity with the truncated ADAMTS13-T7 proteins is obviously higher than that with the overall-length ADAMTS13 proteins; (2) under a denaturation condition, the monoclonal antibody can obviously prevent ADAMTS13 from hydrolyzing VWF , and the suppression effect is enhanced with the increase of the concentration of the monoclonal antibody; (3) the monoclonal antibody disclosed by the invention can be effectively applied to medicine preparation, so that the targeting property and the specificity of a medicine are improved.

This disclosure provides methods and compositions for treating TTP based on transfusion of a relatively small number of genetically modified red blood cells. The genetically modified red blood cells express a fusion protein including a fragment of ADAMTS13 that is enzymatically active against von Willebrand factor (VWF). The fragments of ADAMTS13 can be resistant to the inhibitors, e.g., the auto-immune antibodies, which are responsible for the acquired form of TTP.

The invention relates to an isolated monoclonal antibody that specifically binds to the D4 domain of VWF, competes for binding to VWF D4 domain with ADAMTS13 and partially inhibits ADAMTS 13 -mediated degradation of VWF. More particularly, the invention relates to an isolated monoclonal antibody comprising a heavy chain wherein the variable domain comprises at least one CDR having a sequence selected from the group consisting of SEQ ID NO: 3 for H-CDR1, SEQ ID NO: 4 for H-CDR2 and SEQ ID NO: 5 for H-CDR3 and a light chain wherein the variable domain comprises at least one CDR having a sequence selected from the group consisting of SEQ ID NO: 7 for L-CDR1, SEQ ID NO: 8 for L-CDR2 and SEQ ID NO: 9 for L-CDR3. Antibodies of the invention are presented to be useful in for the prevention or the treatment of bleeding episodes, such as bleeding episodes occurring in patients with aortic stenosis or patients with ventricular assist devices (VAD).

The disclosure provides a method for treating and/or preventing graft rejection with A Disintegrin And Metalloproteinase with Thrombospondin type 1 motif, member-13 (ADAMTS13). The disclosure provides a method for increasing ADAMTS13-mediated von Willebrand factor (VWF) cleavage in a subject that received a graft by administering ADAMTS13. The disclosure also provides a method of determining the likelihood that a subject will rejection a graft.