Impurities in the solution are washed with a washing buffer so that the proteins left on the column are mostly the serine protease having binding affinity with the compound. The serine protease to be purified is retained in the column through binding activity with the compound. A solution comprising the serine protease to be purified is loaded to the affinity column. In some embodiments, the affinity solid support is packed into a column, which is referred to as an affinity column. ![]() The solid support having a small molecule compound bound thereto is referred to as an affinity solid support. The compounds described herein have binding affinity with the serine protease to be purified (e.g., the compounds are ligands of the serine protease), and can be covalently attached to an activated solid support, such as a resin. In some embodiments, the modified fXa protein comprises the amino acid sequence of SEQ ID NO: 2 or a polypeptide having at least about 80% sequence identity to SEQ ID NO: 2. In one aspect, the serine protease comprises a modified derivative of a fXa protein. SUMMARYÄisclosed herein are compounds, compositions, methods and kits for purifying a serine protease. Certain modifications introduced to fXa, however, pose several challenges for purification since routine methods for purification of clotting factors cannot be used for r-Antidote. The modified derivatives of fXa bind to and/or substantially neutralize the anticoagulant. 8,268,783 (which is herein incorporated by reference in its entirety), including r-Antidote, are useful as antidotes to anticoagulants targeting fXa. Previously reported modified derivatives of factor Xa (fXa), such as those described in U.S. For safety considerations, it is also advantageous to have an anticoagulant-antidote pair in the development of new anticoagulant drugs. Thus, specific and effective antidotes to all forms of anticoagulant therapy are highly desirable. ![]() One of the major limitations of anticoagulant therapy, however, is the bleeding risk associated with treatment, and limitations on the ability to rapidly reverse the anticoagulant activity in case of overdosing or if an urgent surgical procedure is required. BACKGROUNDĪnticoagulants serve a need in the marketplace in treatment or prevention of undesired thrombosis in patients with a tendency to form blood clots, such as, for example, those patients having clotting disorders, confined to periods of immobility or undergoing medical surgeries. This disclosure relates to compounds, compositions, methods and kits for the purification of serine proteases, such as factor Xa derivatives. 14, 2013, each of which is hereby incorporated by reference in its entirety. 27, 2012, and is a continuation in part of U.S. This application claims the benefit under 35 U.S.C.
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