33 results about "Pro-urokinase" patented technology

The present invention provides a purification method of recombinant human prourokinase, including protein capture, gel chromatography, cation exchange chromatography, affinity chromatography, anion exchange chromatography, and cation exchange chromatography: the method increases purification steps, Optimize the purification conditions to increase the protein purity to over 98%.

A preparation process for separating and purifying recombinant human pro-urokinase from recombinant Escherichia coli fermentation broth applied in the technical field of biopharmaceuticals, the preparation process comprises the following steps, using the artificially synthesized human pro-urokinase whole gene sequence as a template, by Obtain the target gene fragment of human prourokinase by PCR, construct a prokaryotic expression vector containing human prourokinase, transform the expression vector into Escherichia coli, screen and pick positive clones, which are genetically engineered strains of human prourokinase; The engineered strain of prourokinase was inoculated into solid LB medium, picked a single colony and inoculated into liquid LB medium, cultured on a shaker at 32°C overnight, then inoculated into the fermentation medium in proportion, and cultivated on a shaker at 32°C overnight, Then inoculate into the fermenter according to the proportion, raise the temperature to 42° C. to induce culture for 4 hours, collect the bacteria by centrifugation, disrupt the bacteria by ultrasonic, and collect the precipitate by centrifugation, which is the inclusion body of human prourokinase. The invention is not limited by natural resources, has the advantages of less investment, high output, large profit margin, unique thrombolytic mechanism, obvious effect, and no or low toxicity and side effects.

The invention belongs to the field of pharmacy, more specifically, the invention relates to a medicinal composition containing reactive pro-urokinase, its cryodesiccation method and freeze-dried preparation.

The invention discloses a manufacturing method of an adjustable liver damage animal model and a special DNA fragment thereof; a DNA fragment I is formed by sequentially connecting liver super promoters and antisense tetracycline activator coding genes of a Tet-on gene expression system from upstream to downstream; an DNI fragment II is formed by sequentially connecting tetracycline reaction factors, promoters and prourokinase activator coding genes of the Tet-off / Tet-on gene expression system from upstream to downstream; a recombination expression vector carrying the DNI fragment II is used for converting mammalian cells, the cells are cultivated to obtain recombinant adenovirus. The DNA fragment I is led in the animal to obtain a transgenosis first-construction animal; the transgenosis first-construction animal and scid / bg animal are back-crossed to obtain the animal carrying the DNA fragment I with the scid / bg background; the recombinant adenovirus is used for injecting the animal carrying the DNA fragment I with the scid / bg background, so as to obtain the adjustable liver damage animal model; in the model, the prourokinase activator expression is activated by doxycycline, the expression intensity is changed along with the change of dosage, the specificity is very high.