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Recombinant human platelet derived growth factor fermentation process

A growth factor and platelet technology, which is applied in the field of fermentation of recombinant human platelet-derived growth factor to achieve the effect of low level and favorable for purification

Inactive Publication Date: 2004-06-30
TIANJIN TASLY PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

(The literature comes from High Yield Production from Pichia pastoris Yeast: A Protocol for Benchtop FermentationBy Julia Cino, PhD http: www.nbsc.com / papers / ABL_Pichia.htm) However, according to the report of this literature, expressing foreign proteins generally requires 100-140 peaks in about an hour

Method used

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  • Recombinant human platelet derived growth factor fermentation process
  • Recombinant human platelet derived growth factor fermentation process
  • Recombinant human platelet derived growth factor fermentation process

Examples

Experimental program
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Embodiment 1

[0031] The construction of embodiment 1 engineering strain

[0032] The PDGF-B gene was correctly placed in the yeast expression vector pPICZ-αA by conventional DNA recombination technology, and the yeast strain SMD1168H was transformed, and the engineering strain secreting and expressing rhPDGF-BB was established through screening. Reference books include "Molecular Cloning Technical Guide", "Molecular Biology Technical Guide".

Embodiment 2

[0033] Activation of embodiment 2 engineering strains

[0034] 1) The composition of the plate medium: containing 1% yeast powder (), 2% peptone, 1.6% agar, 2% glucose (freshly added before use).

[0035] 2) Pick a little of the glycerol tube strain stored in the freezer and apply it to the above-mentioned medium, culture at 30°C for 3 days, the colonies are plump, take out and store at 4°C.

Embodiment 3

[0036] The preparation of embodiment 3 primary seeds

[0037] 1) The composition of the medium: 1% yeast powder, 2% peptone, and 2% glucose (freshly added before use).

[0038] 2) Pick the seeds from the plate and inoculate the above-mentioned Erlenmeyer flask culture medium, culture at 30° C., 250 rpm, for 24 hours. OD 600 is 4.0.

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Abstract

The invention relates to a fermenting method to produce reformation human-platelet source growth factor (rhPDGF-BB), including: 1) constructing high-efficiency expression engineering bacterial strain which contains reformation platelet source growth factor destination gene; 2) culturing one-level seed; 3) culturing two-level seed; 4) placing in the cylinder to ferment; 5) collecting supernatant liquid for purifying. It largely shortens the peak-expressing time of the protein.

Description

technical field [0001] The invention relates to a fermentation production method of recombinant human cytokines, in particular, the invention relates to a novel fermentation production method of recombinant human platelet-derived growth factor (rhPDGF-BB). Background technique [0002] Diabetes is one of the most common diseases worldwide. Statistics in 1998 showed that there were approximately 143 million people with diabetes worldwide. There are currently about 16 million people in the United States suffering from diabetes, and the number of diabetics in China is as high as more than 40 million, and it is increasing at a rate of 1‰ every year. According to estimates by WHO experts, by 2025 there will be 300 million diabetic patients worldwide, and 75% of them are in developing countries such as India and China. [0003] A special problem of diabetics is the occurrence of ulcers on the feet and lower extremities, referred to as diabetic foot (dry gangrene). Peripheral ne...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/02
Inventor 卓冰
Owner TIANJIN TASLY PHARMA CO LTD
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