Variable-conformation recombinant interferon crystal, and three-dimensional structure and use thereof

A technology of recombinant interferon and three-dimensional structure, applied in the field of recombinant interferon crystals, can solve the problems of spatial conformation and biological efficacy changes, and achieve the effects of low side effects, high antiviral activity, and reduced toxic and side effects

Inactive Publication Date: 2011-06-22
SICHUAN HUIYANG LIFE SCI & TECH CORP
View PDF8 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The recombinant interferon has the same amino acid sequence as Ganfuj

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
  • Variable-conformation recombinant interferon crystal, and three-dimensional structure and use thereof
  • Variable-conformation recombinant interferon crystal, and three-dimensional structure and use thereof
  • Variable-conformation recombinant interferon crystal, and three-dimensional structure and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0115] Example 1

[0116] Preparation of recombinant interferon rSIFN-co

[0117] This embodiment is a method for preparing recombinant interferon rSIFN-co (hereinafter also referred to as "rSIFN-co") (stock solution) (for details, see Examples 1 and 2 in the specification of U.S. Patent No. 7,364,724 and the specification of Chinese Patent Publication No. CN1740197 Methods described on pages 11-17).

[0118] 1. Gene cloning:

[0119] According to published coding DNA sequence and deduced amino acid sequence data (Klein ML, Bartley TD, Lai PH, et al., Structural characterization of recombinant consensus interferon-alpha. Journal of Chromatography, 1988; 454: 205-215), the large intestine was used Bacillus preferentially express codons (The Wisconsin Package, by Genetics Computer Group, Inc. Copyright 1992, Medison, Wisconsin, USA). Under the condition that the amino acid sequence remains unchanged, the DNA coding sequence is molecularly designed, and then artificially synthesized. ...

Example Embodiment

[0187] Example 2

[0188] Recombinant interferon preparation

[0189] Freeze-dried injection (lyophilized powder) formula

[0190] The rSIFN-co stock solution of the present invention 34.5μg / ml

[0191] pH 7.0 phosphate buffer 10mmol / L

[0192] Glycine 0.4mol / L

[0193] Preparation process: Weigh according to the formula, dissolve in sterile pyrogen-free injection water, filter and sterilize with a 0.22μm pore filter membrane, store at 6-10°C, take a sample for sterility and pyrogen inspection, and pack it into a vial. The single dose of 0.3-0.5 bottle is divided into the lyophilizer and freeze-dried.

[0194] Formula of aqueous injection

[0195] The rSIFN-co stock solution of the present invention 34.5μg / ml

[0196] pH 7.0 phosphate buffer 25mmol / L

[0197] Sodium chloride 0.4mol / L

[0198] Preparation process: Weigh according to the formula, dissolve in sterile, pyrogen-free injection water, filter and sterilize with a 0.22μm pore filter membrane, store at 6-10°C, take a sample for steri...

Example Embodiment

[0199] Example 3

[0200] Crystallization of recombinant interferon

[0201] The preparation of high-quality rSIFN-co protein single crystal is a prerequisite for determining its crystal structure. The rSIFN-co sample used for crystal growth was obtained from the rSIFN-co of the present invention as described above. The method, technical process and crystallization conditions of rSIFN-co single crystal preparation, and its crystallographic parameters are as follows:

[0202] The rSIFN-co freeze-dried powder of the present invention is dissolved in pure water and stored at low temperature (-20°C), and the initial protein concentration is 0.42 mg / ml. The rSIFN-co protein sample was concentrated to 3-3.5mg / ml before crystallization and used for crystal growth experiments immediately. The crystals were cultured at room temperature (293k) using the hanging drop vapor phase diffusion method.

[0203] In the early stage of crystallization research, rSIFN-co crystallites can appear in a v...

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 provides a variable-conformation recombinant interferon crystal, which has the same amino acid sequence as a human interferon and a three-dimensional structure which is different from that of interferon (IFN)-alpha2b. The interferon of the invention has improved bioactivity. The invention also provides a structural model for medicine screening and/or medicine design.

Description

technical field [0001] The present invention mainly relates to a recombinant interferon crystal with changed conformation, its crystallization method, its three-dimensional structure and the application of the crystal and the three-dimensional structure. Background technique [0002] Interferon (IFN) is a soluble protein produced by a variety of cells, which has a variety of important biological functions, including antiviral, antitumor and immunomodulatory functions. Interferon can be divided into type I, type II and type III interferon according to the difference of producing cell type, receptor, biological activity, etc. Type I interferons are mainly induced by viruses and synthetic double-stranded RNA, so they are also called antiviral interferons. There are three types of type I interferons: IFNα, INFβ, and IFNω. Type II interferon, also known as immune interferon or IFNγ, is produced by T cells and is an important immune regulator in the body. Type III interferons a...

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): C07K14/555C07K7/64G01N33/68G06F19/00A61K38/21A61P31/12A61P35/00A61P37/02G16B15/30
CPCG06F19/16C07K2299/00G01N2500/04G06F19/706C07K14/56A61K38/00G16C20/20G16B15/00G16C20/50A61P1/04A61P1/16A61P1/18A61P11/00A61P11/02A61P13/08A61P13/10A61P13/12A61P15/00A61P17/00A61P19/00A61P21/00A61P25/00A61P31/12A61P31/14A61P31/16A61P31/18A61P31/20A61P31/22A61P35/00A61P35/02A61P37/02A61P37/04A61P5/00A61P7/00A61P9/00Y02A50/30Y02A90/10G16B15/30C07K14/555
Inventor 魏光文王大成
Owner SICHUAN HUIYANG LIFE SCI & TECH CORP
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