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Preparation of human SDF-1 alpha, human SDF-1 alpha obtained therefrom and use thereof

A technology of stromal cells and derived factors, applied in the preparation method of peptides, medical preparations containing active ingredients, applications, etc., can solve the problems of high cost, unfavorable large-scale production, and cumbersome steps

Inactive Publication Date: 2008-10-08
再生医药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of using this method to purify recombinant proteins is that the steps are cumbersome and the cost is high
In addition, the yield of protein is not high, and only 400ug of protein can be obtained from 1L of bacterial liquid, so it is not conducive to large-scale production

Method used

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  • Preparation of human SDF-1 alpha, human SDF-1 alpha obtained therefrom and use thereof
  • Preparation of human SDF-1 alpha, human SDF-1 alpha obtained therefrom and use thereof
  • Preparation of human SDF-1 alpha, human SDF-1 alpha obtained therefrom and use thereof

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Experimental program
Comparison scheme
Effect test

Embodiment

[0060] 1. Materials and methods

[0061] 1.1 Experimental materials

[0062] 1.1.1 Expression plasmids and strains

[0063] Escherichia coli expression plasmid pET28a(+) and Escherichia coli BL21(DE3) were purchased from Shanghai Gene Company. Using conventional methods, human bone marrow cDNA was obtained by extracting total RNA from human bone marrow cells and reverse transcription.

[0064] 1.1.2 Animals

[0065] SPF grade male 8-week-old BALB / c mice, weighing 20±1 g, were purchased from Shanghai Slack Experimental Animal Co., Ltd.

[0066] 1.1.3 Related reagents

[0067] DNA restriction enzyme Nco I, Xho I, 10×PCR buffer, dNTP Mix, KOD polymerase, T4 DNA ligase, T4 DNA ligase buffer, D2000 DNA Marker, 6×Loading buffer and 10×Tango buffer purchased from Shanghai Jingmei Biological Company.

[0068] The PCR product purification kit and plasmid extraction kit were purchased from Tianwei Times Company; the chromogenic substrate Limulus kit was purchased from Xiamen Limul...

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Abstract

The invention provides a novel method to prepare a purified human stromal derived factor-1 Alpha(hSDF-1 Alpha), which can greatly simplify purification process, save cost, and improve purity and yield of the produce. The human stromal derived factor-1 Alpha prepared through the method is capable of activating the chemotaxis of T cells and mobilizing medullary hematopoiesis. The invention also provides a new human stromal derived factor-1 Alpha, the pharmaceutical composite and the application thereof.

Description

technical field [0001] The present invention relates to the preparation and purification of human SDF-1α, the human SDF-1α thus prepared and its application. Background technique [0002] The regulatory effect of chemokines on bone marrow hematopoiesis has become the focus of the research and development of a new generation of hematopoietic system drugs. The effects of chemokines on bone marrow hematopoiesis include: promoting bone marrow hematopoiesis and inhibiting hematopoietic cell apoptosis; it is a factor necessary for the generation of platelets in bone marrow, and has a therapeutic effect on thrombocytopenia caused by chemotherapy; it can promote the maintenance of artificial hematopoietic cell expansion and activity in vitro ; Promote the homing, proliferation and maturation of CD34+ hematopoietic stem / progenitor cells (Chai Kexia, Jia Naiyong. Research progress in the regulation of chemokines and their receptors on hematopoietic cells, Clinical Metaphysics, 2003, 1...

Claims

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Application Information

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IPC IPC(8): C07K1/14C07K14/435C12N15/12C12N1/21A61K38/17A61P7/00A61P7/06
Inventor 韩伟俞眉
Owner 再生医药有限公司
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