Polypeptide organic compound and use thereof in hetero transplantation
An organic compound and xenotransplantation technology, applied in the field of organic compounds and polypeptide organic compounds, can solve the problems of hair loss, ineffective hyperacute rejection of xenotransplantation, upper gastrointestinal bleeding, etc., and achieve the effect of avoiding side effects
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Embodiment 1
[0018] Example 1: Using anti-B blood monoclonal antibody as the target protein, phage display technology was used to screen to obtain positive phage clones.
[0019] Phage display technology uses molecular biology technology to clone a set of synthetic oligonucleotide fragments with a certain length of random sequence into a specific expression vector, so that the expression product is displayed on the surface of the phage in the form of a fusion protein. Since the peptide library contains all possible amino acid sequences of the small peptides of this length, each phage surface protein presents one of the peptides, which is convenient for screening; the screened phage can be amplified by bacteria. The phage peptide library is used to infect E. coli, and the random oligonucleotide fragments recombined into the phage are replicated in E. coli and expressed in the coat protein of the phage. Then, the target protein is coated on the ELISA plate. After mixing this phage peptide librar...
Embodiment 2
[0022] Example 2: Stachyose competitive ELISA experiment
[0023] Test method: Coat 100μl of anti-B blood monoclonal antibody in a 96-well ELISA plate at 4°C overnight, block with 5% BSA at 37°C for 2 hours, then wash the plate 3 times with TBS, add 100μl of positive phage and the final concentration respectively The mixture of 100, 20, 4, 0.8, 0.16, 0.032 mM stachyose was reacted for 1 hour, and a positive phage solution without stachyose was set as a blank control. Wash the plate 6 times with TBS containing 0.1% Tween 20, add 100 μl horseradish peroxidase labeled anti-M13 antibody, bind for 1 hour, wash the plate 6 times with TBS containing 0.1% Tween 20, develop color with TMB, and use enzyme The standard instrument measures the absorbance. Calculate the inhibition rate according to the formula: [A 450 (-s)-A 450 (+s)] / A 450 (-s)×100%, A 450 (-s) is the absorbance without stachyose, A 450 (+s) is the absorbance of adding stachyose.
[0024] Test results: Stachyose can effective...
Embodiment 3
[0026] Example 3: Polypeptides inhibit the agglutination reaction between human serum and porcine red blood cells
[0027]Test method: The prepared fresh 1% porcine red blood cells were suspended in phosphate buffer at pH 7.4 for later use. In the wells of the U-type hemagglutination plate, 40μl of peptides were diluted with phosphate buffer sequentially, and then 40μl of appropriately diluted type A human serum and 40μl of 1% porcine red blood cells were added to each well. Then, the mixed solution was shaken with a shaker for 1 minute at room temperature, and then allowed to stand for 1 hour before observing the result of agglutination. If the red blood cells settle to the bottom of the hole as a round dot with a smooth edge, then this is no agglutination. If the red blood cells form a network and do not sink to the bottom of the hole to form a round dot with a smooth edge, this is agglomeration. The test has blank, positive and negative controls.
[0028] Test result: This pept...
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