Specific purified high-molecular urokinase chromatography media as well as preparation method and application thereof

Abstract

A specific purified high-molecular urokinase chromatography media is characterized by having a structure as follows: HOOC-(CH2)4(NH)C-O-R-O-C(NH)NH(CH2)4-CO-X,X=NHC6H5C(NH)NH2, wherein R represents cross-linked agarose substrate. The specific purified high-molecular urokinase chromatography media has the advantages of being simple in preparation process and mild for reaction conditions, being applicable to high-efficiency separation of high-molecular urokinase media and low-molecular urokinase media, and being capable of implementing industrial production of high-purification and high-molecular urokinase.

Description

technical field [0001] The invention relates to a high molecular urokinase chromatographic medium and a preparation method, in particular to a specific purification high molecular urokinase chromatographic medium, a preparation method and an application. Background technique [0002] According to the forecast of Datamonitor, a market research organization, by 2015, the total market value created by global novel antithrombotic drugs will reach about 10 billion US dollars. Among them, the thrombolytic agent plasminogen activators (PAs) class of drugs reached 1.5 billion US dollars. The main plasminogen activators include streptokinase (Streptokinase), urokinase (Urokinase), tissue plasminogen activator (tPA) and so on. With the aging of the world's population, the demand for such drugs will inevitably increase further. As one of the thrombolytic drugs, urokinase can directly promote the inactive plasminogen into active plasmin, and hydrolyze the fibrin that makes up the thro...

AI Technical Summary

Problems solved by technology

The above-mentioned improvement and exploration of urokinase purification medium and process only focused on the final recovery rate and purification multiple of urokinase biological activity, but it was difficult to effectively separate high-molecular-weight urokinase and low-molecular-weight urokinase, so the actual effect was limited

The main reason is that traditional affinity chromatography is based on the "pocket" advanced structure composed of the active site (His204, Asp255 and Ser356) of the β-chain C-terminal serine protease catalytic active domain of the urokinase molecule, while the polymer urokinase and Low-molecular-weight urokinase has this natural high-level structure, so it is often difficult to separate high-molecular-weight urokinase and low-molecular-weight urokinase by affinity chromatography alone, and further separation is required by means of gel filtration chromatography and other steps, which is time-consuming Strenuous effort and loss of urokinase activity

Moreover, human urine protein contains many proteins or enzymes with similar "pocket" structures, such as trypsin and urinary pancreatic kallikrein, etc., and general affinity chromatography also lacks resolution for these biochemical molecules