Preparation method for human thymosin beta 4 two-string body protein

A technology of thymosin and protein, applied in the field of medicine and biology, can solve the problems of high price, limited application, complicated process, etc.

Active Publication Date: 2013-08-21
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Thymosin β4 currently used in laboratories and clinical trials is mostly chemically synthesized and expensive; however, the extraction of Tβ4 from mammalian tissues is complicated and the yield is low, which limits its application. How to obtain it in a more economical way This peptide appears to be very important

Method used

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  • Preparation method for human thymosin beta 4 two-string body protein
  • Preparation method for human thymosin beta 4 two-string body protein
  • Preparation method for human thymosin beta 4 two-string body protein

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Embodiment Construction

[0035] The present invention will be described in detail through specific examples below.

[0036] This invention comprises the following steps successively:

[0037] 1. Human Thymosin β 4 Bacterial cell destruction by dimerin

[0038] Induced Culture of Engineering Bacteria BL21 / pET-22b(+)-Tβ Containing Recombinant Human Thymosin β4 Dimer Protein 4 ②, the inducer is a low concentration of IPTG (0.01mM), and the cells are obtained by centrifugation.

[0039] According to the ratio of 1g of bacteria plus 10ml of STE lysis buffer (20mM Tris-HCl pH 8.0, 100mMNaCl, 1mM EDTA), resuspend the bacteria, about 15g, and ultrasonically lyse the bacteria in an ice bath with a power of 300W and a working time of 5s. The interval time is 10s, the whole time is 30min, centrifuge (12000rpm×20min, 4°C), and collect the supernatant.

[0040] 2. Human Thymosin β 4 Purification of dimerized proteins

[0041] Dissolve the supernatant (about 150ml) obtained in step 1 in 20mM Tris-HCl buffer c...

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Abstract

The invention belongs to the technical field of medical biology, and particularly relates to a preparation method for a human thymosin beta 4 two-string body protein. A target protein with purity more than 98% can be obtained through the preparation method, the obtained human thymosin beta 4 two-string body protein has biological activity, the activity of the protein is determined through proliferation experiments and migration experiments, and results show that the human thymosin beta 4 two-string body protein can promote proliferation and migration of cells and the activity of the human thymosin beta 4 two-string body protein is superior to that of a chemical synthesis monomer. The method only uses one chromatographic column to complete separation and purification of the target protein,is simple in steps, rapid and low in cost and lays a foundation for further study and wide application of human thymosin beta 4.

Description

technical field [0001] The invention belongs to the technical field of medicine and biology, and in particular relates to a preparation process of recombinant human thymosin β4 dimer target protein. Background technique [0002] Thymosin β4 is a polypeptide composed of 43 amino acid residues, which is highly conserved in mammals. Unlike prothymosin or thymosinoids, thymosin β4 is a cytoplasmic protein rather than a nuclear protein, and it contains only fewer hydrophobic amino acids and does not contain the Lys-Lys-Xaa-Lys structural region. Its amino acid sequence is as follows: AcSDKPDMAEIEKFDKSKLKKTETQEKNPLPSKETIEQEKQAGES. Thymosin β4 is widely distributed in various tissues, organs and cells, and is closely related to immune function, nerve development and actin function. It can also resist inflammation, promote angiogenesis, wound healing, hair follicle development, and repair damaged myocardium. [0003] Although thymosin β4 has broad application prospects, it is diff...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/02C07K1/20C12R1/19
Inventor 张英起徐天娇李萌张伟王增禄
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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