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17 peptide organic compound and its application

An organic compound and reaction technology, applied in the field of polypeptide organic compounds and organic compounds, can solve problems such as blood transfusion reactions, achieve the effect of avoiding side effects and inhibiting hyperacute rejection

Inactive Publication Date: 2003-04-23
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] One of the objectives of the present invention is to overcome the problems of hyperacute rejection and transfusion reactions caused by incompatibility of blood types in clinical organ transplantation, and to design and provide a 17-peptide organic compound

Method used

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  • 17 peptide organic compound and its application
  • 17 peptide organic compound and its application
  • 17 peptide organic compound and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Using the anti-B blood monoclonal antibody as the target protein, a positive phage clone was obtained by the method of phage display.

[0021] The phage display method uses genetic engineering to clone a set of synthetic oligonucleotide fragments of a certain length and random sequence into a specific expression vector, so that the expression product is presented on the surface of filamentous phage in the form of fusion protein. Since the peptide library contains all possible amino acid sequences of small peptides of this length or most of them, each phage presents one of the peptides, so the library has a large capacity and is easy to screen and amplify. Infect E. coli with the phage peptide library, and the random oligonucleotide fragments recombined into the phage are replicated in E. coli and expressed in the coat protein of the phage. Then, coat the target protein on the microtiter plate. After mixing the phage peptide library with the target protein, w...

Embodiment 2

[0024] Embodiment 2: Competitive ELISA (melibiose) test

[0025] Test method: Coat 96-well ELISA plate with 100 μL / well of anti-B blood monoclonal antibody (titer 1:128), overnight at 4°C. 5% BSA was blocked at room temperature for 2 hours, and the plate was washed 3 times with 5 mM TBS buffer solution. The ELISA solution added was 100 μL of a mixture of different concentrations of melibiose and phage stock solution. The following steps are the same as before. Calculated inhibition rate=(OD450 without melibiose-OD450 with melibiose) / OD450*100% without melibiose

[0026] Test results: Melibiose can very well inhibit the combination of anti-B blood monoclonal antibodies and positive clones, and the effect can be seen at a concentration as low as 0.003mM. With the increase of the concentration of melibiose, the concentration of 300mM The inhibition rate has reached more than 80%, which shows that the positive clones are likely to bind to the site where melibiose binds to the an...

Embodiment 3

[0028] Example 3: Small peptide inhibits the agglutination reaction between type A human serum and pig erythrocytes

[0029] Test method: Suspend the prepared fresh 2% porcine red blood cells in the phosphate buffer solution with pH=7.4 for later use. In the wells of the V-type hemagglutination plate, after 40ul of small peptides are sequentially diluted well by well with phosphate buffer, 40ul of appropriately diluted blood type A human serum and 40ul of 2% porcine red blood cells are added to each well. Then shake the 120ul mixture with a shaker for 1 minute at room temperature, and then let it stand for 1 hour to observe the agglutination result. If the red blood cells settle to the bottom of the well as a round spot with smooth edges, this is not agglutinated. If the red blood cells do not sink to the bottom of the well, but appear as a network with rough edges, this is called agglutination. A blank and a positive control must be set up in the test.

[0030] Test result...

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PUM

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Abstract

The present invention provides a 17 peptide organic compound, it can make specific combination with anti-type B monoclonal antibody, dissolve in water, between all the amino acids they are connected by means of peptide bond. Because it can be specifically combined with anti-type B antibody, it can possess the action of closing anti-B monoclonal antibody so as to prevent the side effect resulted from that the specificity of action site is not strong, it can be used for controlling transfusion reaction resulted from incompatible transfusion. Said invention also can be specifically combined withinternal prestored antibody of anti-pig antigen so as to close its action, and can inhibit the production of pig's heteroplastic transplantation hyperacute rejection reaction.

Description

technical field [0001] The invention belongs to organic compounds, and mainly relates to polypeptide organic compounds, especially 17-peptide organic compounds obtained by artificial chemical synthesis or genetic engineering and their application in the field of medicine. technical background [0002] There are three types of rejection after organ transplantation: hyperacute, acute, and chronic. Hyperacute rejection (HAR) is immunologically due to recognition and binding between antigens on the cell membrane of the transplanted organ and circulating antibodies of the recipient. Because these antibodies are pre-existing in the recipient, hyperacute rejection occurs rapidly after transplantation, usually within minutes or hours after transplantation. Due to the shortage of human organs, the use of pig organs to replace human organs is a hot research topic at present. However, because the pig cell surface antigen Gal-α-1, 3-Gal can combine with the pre-existing natural antibo...

Claims

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Application Information

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IPC IPC(8): A61K38/04A61P37/06A61P43/00C07K7/04C07K7/08
Inventor 詹金彪张晖
Owner ZHEJIANG UNIV
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