Protein compound, and preparation method and application thereof

A complex and protein technology, applied in the field of molecular biology, can solve the problems of high price and short half-life

Pending Publication Date: 2021-03-19
TSINGHUA UNIV
View PDF2 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, human interleukin-1 receptor antagonist (anakinra) has been successfully used in the treatment of rheumatoid arthritis and acute gouty arthritis, but due to its short half-life in vivo, patients need repeated administration to achieve effective, and expensive

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
  • Protein compound, and preparation method and application thereof
  • Protein compound, and preparation method and application thereof
  • Protein compound, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0128] The preparation of embodiment 1 recombinant protein

[0129] In this example, the inventors designed the following 10 recombinant protein sequences based on (VPGKG) repeated polypeptide sequence and human interleukin 1 receptor antagonist (IL-1RA) sequence. For specific sequence information, see Table 2:

[0130] Table 2 Recombinant protein sequence characteristics

[0131]

[0132] 1.1 Obtaining strains expressing recombinant proteins

[0133] The nucleotide sequences corresponding to the above 10 recombinant proteins were synthesized by Sangon Biotech (Shanghai) Co., Ltd. and constructed on the pET25b expression plasmid vector for transformation into prokaryotic expression competent BLR (DE3) of Escherichia coli. The specific embodiment is as follows: add 100 ng of pET25b-A1 (or A2~A10) plasmid into 100 μL of BLR Escherichia coli competent (purchased from Novagen) in an ice bath environment, and maintain it in an ice bath environment for 30 min; then The mixture ...

Embodiment 2

[0138] The preparation of embodiment 2 protein complexes

[0139] Protein complex 1: The purified recombinant protein A1 was mixed with carboxylated PEG at a molar ratio of 1:18, and stirred at 4°C for 1 hour. The mixture was loaded on a desalting chromatography column (purchased from GE), and eluted with 50 mM PB buffer with a pH of 7.2-7.4 to collect protein complex components.

[0140] Protein complex 2: The purified recombinant protein A2 was mixed with carboxylated PEG at a molar ratio of 1:72, and stirred at 4°C for 1 hour. The mixture was loaded on a desalting chromatography column (purchased from GE), and eluted with 50 mM PB buffer with a pH of 7.2-7.4 to collect protein complex components.

[0141] Protein complex 3: The purified recombinant protein A3 was mixed with carboxylated PEG at a molar ratio of 1:288, and stirred at 20-25°C for 4 hours. The mixture was loaded on a desalting chromatography column (purchased from GE), and eluted with 50 mM PB buffer with a p...

Embodiment 3

[0159] Example 3: Determination of protein complex particle size

[0160] The protein complex purified in Example 2 was detected by transmission electron microscopy, and the average particle size of the protein was analyzed. see results Figure 2~3 , figure 2 Indicates electron microscope pictures of protein complexes. image 3 Indicates the average particle size analysis results of proteins.

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

No PUM Login to view more

Abstract

The invention relates to the technical field of molecular biology, and in particular relates to a recombinant protein, a protein compound containing the recombinant protein and application. The recombinant protein disclosed by the invention is composed of a human interleukin 1 receptor antagonist protein and a protein containing a VPGKG repetitive unit, can be used for adjuvant therapy of rheumatoid arthritis, acute gouty arthritis and other rheumatic immune diseases, hereditary diseases and tumours with increased interleukin 1 (interleukin, IL-1), and has a remarkable curative effect.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a protein complex and its preparation method and application. Background technique [0002] Rheumatoid arthritis is a human immune disease, and the exploration of the etiology of the disease is still in its infancy. In addition to the patient's own genetic factors, infection, abnormal immune system, environmental factors (such as exposure to silicon dioxide, infectious agents, vitamin D deficiency in the body), bad living habits (such as smoking, obesity), changes in the microbiome in the body, etc. lead to an increased risk of the disease. Patients with rheumatoid arthritis, the incidence of hypertension, diabetes, hyperlipidemia and obesity were 18.6%, 6.0%, 9.9% and 4.4%, these secondary cardiovascular diseases are the cause A common cause of premature death in patients. Since the disease has little effect on the patients in the early stage of the disease, and the...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/70A61K38/17A61K47/64A61K45/06A61P29/00A61P19/02A61P19/06A61P37/02A61P35/00
CPCC07K14/4703C07K14/00C12N15/70A61K47/64A61K45/06A61P29/00A61P19/02A61P19/06A61P37/02A61P35/00A61K38/00
Inventor 刘凯张洪杰
Owner TSINGHUA UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap