Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Simple and convenient chemical industrial technology for acidic fibroblast growth factor (aFGF)

A fibroblast and growth factor technology, applied in the direction of fibroblast growth factor, growth factor/inducer, recombinant DNA technology, etc., can solve the problems of reduced economic significance, low yield, difficult extraction, etc.

Inactive Publication Date: 2012-02-08
ABZYME BIOTECH
View PDF1 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since haFGF is an endogenous trace active substance in the human body, it is difficult to extract from body fluids or tissues, so the yield is not high, and the economic significance is also greatly reduced

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Simple and convenient chemical industrial technology for acidic fibroblast growth factor (aFGF)
  • Simple and convenient chemical industrial technology for acidic fibroblast growth factor (aFGF)
  • Simple and convenient chemical industrial technology for acidic fibroblast growth factor (aFGF)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0012] 1. Prokaryotic expression of acidic fibroblast growth factor

[0013] The human-derived aFGF gene was connected to the expression vector pET-26b through NdeI and EcoRI restriction sites. The plasmid was subsequently transformed into E. coli BL21(DE3).

[0014] Pick a single clone and insert it into a medium containing 10 mL LB and a final concentration of 100 μg / ml kanamycin, and culture it on a shaker at 37° C. and 250 rpm for about 16 hours.

[0015] Put the above-mentioned bacterial solution into the medium containing 1000mL LB and the final concentration of 100ug / ml kanamycin at a ratio of 1:100, cultivate it on a shaking table at 37°C and 250rpm until the OD600 is about 0.8, and place the bacterial solution in Chill on ice. Add IPTG to a final concentration of 0.8mM, harvest the bacteria after induction at 30°C for 4 hours, centrifuge at 5000rpm for 20min, discard the supernatant, and store the bacteria at -80°C. SDS-PAGE electrophoresis analysis results of aFGF...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Theoretical molecular weightaaaaaaaaaa
Login to View More

Abstract

The invention relates to a simple and convenient chemical industrial technology for prokaryotic expression and purification of humanized acidic fibroblast growth factor (aFGF), and aims to construct engineering bacteria of recombinant haFGF (Human Acidic Fibroblast Growth Factor) gene by using a recombinant DNA (Deoxyribonucleic Acid) technology, realize expression of a large amount of humanized FGF-1 in prokaryotic cells and obtain a target gene with higher purity through simple purification steps. haFGF is produced by using the biological engineering method, so that the technology has the advantages of easiness in forming scale production capacity, high yield and low cost.

Description

technical field [0001] The invention relates to a simplified method for prokaryotic expression and purification of human-sourced acidic fibroblast growth factor (aFGF). Background technique [0002] Fibroblast Growth Factor (FGF for short) is a protein family, and 9 members have been discovered. Their common feature is that they bind to heparin, so they are also called heparin-bound growth factors. Acidic fibroblast growth factor (aFGF, also known as AFGF) is an important member of this family. It is a kind of polypeptide that has nutritional and mitogenic differentiation effects on various cells derived from mesoderm and neuroectoderm. It was originally isolated and purified from bovine brain by Thomas in 1984. In 1986, Michael Jaye cloned the human acidic fibroblast growth factor (haFGF) gene from human brain for the first time, and determined the nucleotide sequence, thus the research on aFGF entered the stage of DNA recombination. [0003] Studies have shown that haFG...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N15/70C07K14/50C07K1/18C07K1/16C12R1/19
Inventor 柴笑梅朱月珍杨宣武吴咪佳
Owner ABZYME BIOTECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com