Structures and application of bifunctional protein and derivatives of bifunctional protein

A kind of use, protein technology, applied in the field of bioengineering, can solve the problems of linker length, immunogenicity toxicity, clinical treatment midway stop, etc.

Active Publication Date: 2010-12-08
CHINA PHARM UNIV
View PDF6 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

(2) Secondly, also due to long-term repeated administration, the immunogenicity and antigenicity of drug molecules have to be considered. If the immunogenicity is too strong, once endogenous antibodies are produced, it will lead to failure of clinical treatment. stop halfway
However, in our previous research

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
  • Structures and application of bifunctional protein and derivatives of bifunctional protein
  • Structures and application of bifunctional protein and derivatives of bifunctional protein
  • Structures and application of bifunctional protein and derivatives of bifunctional protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Molecular construction and sample preparation of recombinant proteins

[0067] The amino acid sequence of the recombinant protein is:

[0068] His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp- Leu-Val-Lys-Gly-Arg-Ala-Met-Gly-Ala-Tyr-Val-Tyr-Arg-Ser-Ala-Phe-Ser-Val-Gly-Leu-Glu-Thr-Tyr-Val-Thr- Ile-Pro-Asn-Met-Pro-Ile-Arg-Phe-Thr-Lys-Ile-Phe-Tyr-Asn-Gln-Gln-Asn-His-Tyr-Asp-Gly-Ser-Thr-Gly-Lys- Phe-His-Cys-Asn-Ile-Pro-Gly-Leu-Tyr-Tyr-Phe-Ala-Tyr-His-Ile-Thr-Val-Tyr-Met-Lys-Asp-Val-Lys-Val-Ser- Leu-Phe-Lys-Lys-Asp-Lys-Ala-Met-Leu-Phe-Thr-Tyr-Asp-Gln-Tyr-Gln-Glu-Asn-Asn-Val-Asp-Gln-Ala-Ser-Gly- Ser-Val-Leu-Leu-His-Leu-Glu-Val-Gly-Asp-Gln-Val-Trp-Leu-Gln-Val-Tyr-Gly-Glu-Gly-Glu-Arg-Asn-Gly-Leu- Tyr-Ala-Asp-Asn-Asp-Asn-Asp-Ser-Thr-Phe-Thr-Gly-Phe-Leu-Leu-Tyr-His-Asp-Thr-Asn

[0069] The recombinant protein has a total composition of 170 amino acids and a theoretical molecular weight of 19430.6 Daltons. According to the c...

Embodiment 2

[0078] Construction and preparation of recombinant proteins

[0079] The amino acid sequence of the recombinant protein is:

[0080] His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln-Met-Glu-Glu-Glu-Ala-Val-Arg-Leu-Phe-Ile-Glu-Trp- Leu-Lys-Asn-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-Gly-Met-Gly-Ala-Tyr-Val-Tyr-Arg-Ser-Ala-Phe- Ser-Val-Gly-Leu-Glu-Thr-Tyr-Val-Thr-Ile-Pro-Asn-Met-Pro-Ile-Arg-Phe-Thr-Lys-Ile-Phe-Tyr-Asn-Gln-Gln- Asn-His-Tyr-Asp-Gly-Ser-Thr-Gly-Lys-Phe-His-Cys-Asn-Ile-Pro-Gly-Leu-Tyr-Tyr-Phe-Ala-Tyr-His-Ile-Thr- Val-Tyr-Met-Lys-Asp-Val-Lys-Val-Ser-Leu-Phe-Lys-Lys-Asp-Lys-Ala-Met-Leu-Phe-Thr-Tyr-Asp-Gln-Tyr-Gln- Glu-Asn-Asn-Val-Asp-Gln-Ala-Ser-Gly-Ser-Val-Leu-Leu-His-Leu-Glu-Val-Gly-Asp-Gln-Val-Trp-Leu-Gln-Val- Tyr-Gly-Glu-Gly-Glu-Arg-Asn-Gly-Leu-Tyr-Ala-Asp-Asn-Asp-Asn-Asp-Ser-Thr-Phe-Thr-Gly-Phe-Leu-Leu-Tyr- His-Asp-Thr-Asn

[0081] The recombinant protein consists of 179 amino acids. According to the codon table of Escherichia coli and combined with...

Embodiment 3

[0088] The polyethylene glycol modification of the recombinant protein obtained in embodiment 1 and the preparation of the modified product

[0089] Accurately measure 10 mg (1 mg / ml, 10 ml) of the recombinant protein obtained in Example 1, and dialyze to replace the buffer system. After dialysis, the final reaction system was BufferR5 (100mMNaCl, 5mM EDTA, 20mMNa 2 HPO 4 -NaH 2 PO 4 , pH8.0). Add maleimide-activated polyethylene glycol (molecular weight 20,000 Daltons) to the reaction system at a molar ratio of 1:10, place the reaction mixture in a 4-degree thermostat for stirring reaction, and the rotating speed of the stirring device is set at 60 rpm. After 24 hours of reaction, the reaction mixture was diluted 1:5 (V / V) to Buffer D5 (5 mM EDTA, 20 mM Tris-HCl, pH 8.0) and purified by liquid chromatography.

[0090] The liquid phase system adopts GE Healthcare's AKTA medium and low pressure device, the chromatographic column is a 20×1.6 (I.D., cm) silanized glass colu...

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 structures and application of a bifunctional protein and a derivative of the bifunctional protein and belongs to the technical field of biological engineering. The technology of computer aided molecular design is adopted and the means of joint engineering are combined to design and prepare a recombinant protein with double functions of reducing blood sugar and blood fat; and on this basis, the obtained target recombinant protein is subjected to polyethylene glycol chemical modification. The target recombinant protein and a chemically modified derivative thereof have obvious blood sugar and blood fat reducing activities for mice models with diabetes. The molecule of the protein can be used for treating diabetes and concurrent diseases of diabetes.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a molecular structure of a bifunctional protein and its derivatives, and the use of the molecule with the structure in the treatment of diabetes and its concurrent diseases. Background technique [0002] Diabetes mellitus (DM) is a metabolic system disease with multiple etiologies, characterized by chronic hyperglycemia accompanied by disorders of sugar, fat and protein metabolism caused by defects in insulin secretion and / or action. The disease can be divided into two types: type 1 diabetes, also known as insulin-dependent diabetes; type 2 diabetes (type2diabetesmellitus, T2DM), also known as non-insulin-dependent diabetes. Among them, T2DM accounts for more than 95% of the total incidence. [0003] With the improvement of living standards and the aggravation of aging, the number of diabetic patients in my country is increasing. According to authoritative data, by 2030, t...

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): C07K14/47C07K1/107A61K38/17A61K47/48A61P3/10A61P9/10A61P3/04A61P25/28A61K47/64
Inventor 姚文兵高明明高向东马辰
Owner CHINA PHARM 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