Antifibrin-2 antibody sequence, tetrapeptide chain molecule and immunoglobulin molecule
A fibrin and sequence technology, applied in the direction of antiviral immunoglobulin, immunoglobulin, peptide, etc.
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Embodiment 1
[0063] Example 1: Construction of chicken-derived antiserum type 4 avian adenovirus fibrin-2 single-chain antibody library
[0064] 1. Isolation of Chicken Peripheral Blood Lymphocytes
[0065] According to the International Animal Immunization Manual, 5 experimental chickens were immunized with serotype 4 avian adenovirus inactivated vaccine. Each immunization interval was 2 weeks, and a total of 3 immunizations were performed. Two weeks after the third immunization, the peripheral blood of the chickens was collected. Take 5-10mL of fresh blood and mix evenly with whole blood and tissue diluent at a ratio of 1:1-1:2. Add cell separation solution to a 15mL centrifuge tube, and gently add an equal volume of diluted anticoagulant blood along the tube wall. The horizontal centrifuge is centrifuged at a speed of 400g-800g and a time of 15min-25min. After centrifugation, the liquid in the centrifuge tube should be divided into four layers, from top to bottom: the first layer: pl...
Embodiment 2
[0072] Example 2: Screening of chicken-derived antiserum type 4 avian adenovirus fibrin-2 single-chain antibody library
[0073] 1. Preparation of Phage Antibody
[0074] Add 1 mL of phage primary antibody library to 3 mL of XLI-Blue bacterial solution with OD600=0.5, place at 37°C for 45 min, add 6 mL of LB liquid medium containing Amp+ (100 μg / mL), and add glucose (1 mol / mL) at a ratio of 1:1000 L) Continue to cultivate, when the OD600 of the bacterial solution is 0.5, add the helper phage M13K07, place it in a 37°C incubator for 30min, and then incubate at 220rpm / min at 37°C for 1h, centrifuge the bacterial solution at 3500rpm for 10min, discard the supernatant, and use an equal volume LB liquid medium containing Amp+ (100 μg / mL) and Kana (50 μg / mL) was added with IPTG (1 mol / L) at a ratio of 1:1000, 220 rpm / min, 37°C, and shaken for 6 hours. Centrifuge the culture medium at 12000rpm for 10-20min, and collect the supernatant. Add PEG8000 at a ratio of 1:4-1:5, invert and ...
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