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.

The present invention relates to formulations of ADAMTS13 with enhanced or desirable properties. As such, the invention provides liquid and lyophilized formulations of ADAMTS13 that are suitable for pharmaceutical administration. Among other aspects, the present invention also provides methods of treating various diseases and conditions related to VWF and/or ADAMTS13 dysfunction in a subject. Also provided herein are kits comprising ADAMTS13 formulations useful for the treatment of various diseases and conditions.

Compositions and methods for the treatment of thrombotic thrombocytopenic purpura are disclosed.

Among other aspects, the present invention relates to cell culture conditions for producing high molecular weight vWF, in particular, highly multimericWF with a high specific activity and ADAMTS13 with a high specific activity. The cell culture conditions of the present invention can include, for example, a cell culture medium with an increased copper concentration and/or cell culture supernatant with a low ammonium (NH.sub.4.sup.+) concentration. The present invention also provides methods for cultivating cells in the cell culture conditions to express high molecular weight vWF and rA13 having high specific activities.

Provided herein are methods and compositions for increasing production of a target protein or functional RNA by a cell.

A pharmaceutical formulation of plasma protein, specifically ADAMTS-13 protein, and a composition for preventing or treating thrombotic disease containing the same are disclosed. The pharmaceutical formulation significantly improves the stability of plasma protein whose pharmacological activity and quality deteriorates during long term storage due to its high risk of being contaminated and denatured immediately after separation and purification from blood. The formulation is capable of maintaining the colloidal stability, refrigeration stability, purity, and inhibition of aggregation of the plasma protein at high levels, and allows the cake appearance after lyophilization to be maintained well for a long period of time.

A protease-resistant ADAMTS13 mutant protein or nucleic acid encoding the ADAMTS13 mutant protein is provided. The ADAMTS13 mutant protein comprises a mammalian ADAMTS13 protein in which one or more protease cleavage sites within the protein are replaced with amino acid sequence that is resistant to protease cleavage, and the mutant protein retains von Willebrand factor (VWF)-cleaving activity. The protease-resistant ADAMTS13 mutant is useful as a thrombolytic agent to treat common thrombotic disorders, including stroke, myocardial infarction, venous thromboembolism, and rare microvascular thrombotic disorders like thrombotic thrombocytopenia purpura (TTP).

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.

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 disclosure provides a method for treating sepsis caused by a microorganism infection. The disclosure also relates to a method for identifying and discovering compounds useful in treating sepsis as well as a method for determining appropriate approaches for treating sepsis. In particular, the methods of the disclosure are based on the discovery by the present inventor that sepsis is caused by exotoxin(s) of microorganism or by constitutive proteins that function as exotoxins as in the case of viruses.