Monoclonal antibody against RBD region of novel coronavirus SARS-CoV-2 spike protein and application of monoclonal antibody
A monoclonal antibody, spike protein technology, applied in the direction of antiviral immunoglobulin, application, antibody, etc., can solve problems such as toxic side effects, no obvious therapeutic effect, etc., to achieve strong specificity, prevention or/and treatment of new coronavirus Pneumonia, the effect of high expression
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[0034] 1. Amplification of phage display antibody library:
[0035] Inoculate 500 μL of Escherichia coli TG-1 containing Tomlinson I+J phage display antibody library (MRC HGMP Resource Center, UK) into 25 mL of 2YT medium (1.6% Tryptone, 1% Yeast Extract) containing 100 μg / mL ampicillin and 1% glucose , 0.5% NaCl), cultured at 37°C to OD 600 is 0.4, add 10 9cfu (Colony FormationUnit) KM13 helper phage (MRC HGMP Resource Center, UK), after infection for 1 hour at 37°C, centrifuge at 3000g for 30 minutes, discard the supernatant, and use bacteriophage containing 100μg / ml ampicillin, 50μg / ml kanamycin and 0.1% Glucose 2YT medium 50mL (1.6% Tryptone, 1% Yeast Extract, 0.5% NaCl) suspended bacteria, 30°C, 250rpm speed shake for 16 hours, the next day, 5000g, 30 minutes of centrifugation of the culture medium, separation and recovery of supernatant 40mL , add 10mL of PEG / NaCl solution to the supernatant, mix well, place on ice for 30 minutes, centrifuge at 5000g for 30 minutes, di...
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