Renaturation separation purification method of tissue plasminogen activator mutant

A technology of plasminogen and activator, which is applied in the field of separation and purification of recombinant protein in medical biotechnology, can solve problems such as high cost, and achieve the effects of reducing loss, saving time and reducing production cost.
CN1556206AInactive Publication Date: 2004-12-22CHINA PHARM UNIV
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
CHINA PHARM UNIV
Publication Date
2004-12-22
Estimated Expiration
Not applicable · inactive patent

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Abstract

A process for renaturating, separating and purifying the mutant of tissue plasminogen activator features that the imidazole and urea are used to make the tissue plasminogen activator 'Ruitipu enzyme' be correctly folded in order to have activity and make it be renaturated, separated and purified simultaneously.
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Description

1. Technical field

[0001] The invention belongs to the technical field of separation and purification of recombinant protein of medical biotechnology. 2. Background technology

[0002] Reteplase is a gene recombinant human tissue plasminogen activator mutant expressed in Escherichia coli by site-directed mutagenesis technology. It is a single-chain aglycosylated polypeptide, containing 355 amino acids, and has the 1-3, 176-527 amino acid coding sequence of natural human tissue plasminogen activator (tissue plasminogen activator, t-PA). Reteplase, which only carries the kringle2 domain and protease domain of wild-type t-PA, has become a comparative therapy because of its long half-life of 16-20 minutes, faster and more thorough thrombolysis and less risk of intracranial hemorrhage. Ideal thrombolytic drug.

[0003] Since reteplase contains 10 pairs of disulfide bonds, it forms inclusion bodies when expressed in Escherichia coli, and it is difficult to correctly fold and ref...

Claims

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