Avian-derived anti-avian-flu neuraminidase monoclonal antibody, preparation method therefor and application of avian-derived anti-avian-flu neuraminidase monoclonal antibody
A kind of monoclonal antibody, single amino acid enzyme technology, applied in the field of biomedicine
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Embodiment 1
[0029] A preparation method of chicken-derived anti-avian influenza neuraminidase monoclonal antibody, comprising the following steps:
[0030] (1) Extract and amplify the full-length gene of the light chain of the monoclonal antibody from the start codon of the signal peptide to the stop codon from the hybridoma cells secreting the mouse monoclonal antibody and clone it into a mammalian expression vector to obtain A plasmid expressing the light chain;
[0031] The mammalian expression vectors include pCDNA3, pCDNA3.1, pCMV, pCAGGS, pIRES.
[0032] (2) Extract and amplify the Fd fragment sequence from the start codon of the signal peptide to the end of the CH1 region of the heavy chain chain of the monoclonal antibody from the hybridoma cells secreting the mouse monoclonal antibody, and then extract the amplified sequence from the spleen of the immunized chicken The gene sequence of the Fc fragment from the start amino acid codon to the stop codon of the chicken IgY antibody ...
Embodiment 2
[0036] 1. Construction of vector pCAGGS-LC expressing antibody light chain
[0037] RNA is routinely extracted from hybridoma cells secreting mouse monoclonal antibodies and reverse-transcribed into cDNA. PCR amplifies the full-length gene of the light chain of the monoclonal antibody from the signal peptide start codon to the stop codon and utilizes Sac I and NheI restriction sites were cloned into the mammalian expression vector pCAGGS to obtain the pCAGGS-LC plasmid expressing the light chain (see figure 1 ).
[0038] 2. Construction of expression recombinant heavy chain vector pCAGGS-FdchFc
[0039] PCR amplifies the Fd fragment sequence from the start codon of the signal peptide to the end of the CH1 region of the heavy chain of the monoclonal antibody. RNA was extracted from the spleen of adult immunized chickens and reverse-transcribed into cDNA, and the Fc fragment gene sequence of the chicken IgY antibody from the N-terminal start amino acid codon to the stop codon ...
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