Blood group B epitope mimic peptide and application thereof
An epitope mimic peptide and blood group antigen technology, applied in the field of biochemistry, can solve the problems of difficult modification, complicated B blood group sugar antigen synthesis technology, and high cost
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example 1B
[0020] Discovery and Screening of Example 1B Blood Group Antigen Epitope Mimetic Peptides
[0021] The B blood group antigen epitope mimetic peptide of the present invention is initially screened out from a phage random 12 peptide library, and the specific method is as follows:
[0022] 1. The titer determination of the phage random 12-peptide library ensures that the amount of phage input is about 1.5×10 11 pfu.
[0023] 2. Affinity panning method of phage random peptide library
[0024] (1) Coat 100 mg / L, 100 μl / well anti-blood group B antigen monoclonal antibody on an ELISA plate, shake gently in a wet box, and incubate overnight at 4°C. Take 10 μl of the peptide library stock solution and dilute it to 100 μl with TBS, add it to the wells of the ELISA plate, 100 μl / well, shake slowly at room temperature for 1 hour. Plates were washed 10 times with 0.1% TBST. Add 100 μl of 0.2M glycine-hydrochloric acid (pH2.2), dilute and amplify the eluate with LB medium, and measure t...
example 2
[0031] Example 2 Study on P / F-pIRES, P / F-M-pIRES Recombinant Plasmids and Their Antitumor Effects
[0032] 1. Construction of P / F-pIRES and P / F-M-pIRES recombinant plasmids
[0033] ①. Construction of P / F-pIRES recombinant plasmid
[0034] Using the sequence of SEQ NO.2 as a template, using dATP, dGTP, dCTP and dTTP as raw materials, through the steps of deprotection group activation, ligation, closure, oxidation and other steps to form a transmembrane mimetic peptide and FAS gene in ABI 394 DNA / RNA synthesizer The P / F fusion gene fused with region and intracellular segment, the upstream of the fusion gene contains the Xho I restriction site, and the downstream contains the Mlu I restriction site. The P / Fas fusion gene and pIRES plasmid were double digested with Xho I and Mlu I. The digestion reaction system was 40 μL, including: 8 μL 10×Tango Buffer, 2 μL Xho I, 2 μL Mlu I, 10 μL DNA and 18 μL sterile water. The reaction system was vortexed and mixed, then placed in a PCR ...
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