Hybridoma cell capable of secreting anti-Ad3FK monoclonal antibody, monoclonal antibody and preparation method and application thereof
A monoclonal antibody, hybridoma cell technology, applied in the biological field, can solve the problem of no specific and effective therapeutic drugs
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[0041] One embodiment of the method for preparing hybridoma cells that can secrete anti-Ad3FK monoclonal antibody includes the following steps S110-S170.
[0042] S110. Prepare Ad3FK antigen.
[0043] Specifically, the operation of preparing the Ad3FK antigen includes S111-S117.
[0044] S111. Codon-optimizing the gene of the head domain (fiber knob) of human adenovirus type 3 ciliary protein, so as to adapt to expression in Escherichia coli.
[0045] Further, as shown in SEQ ID No: 5 of the gene sequence of the head domain (fiber knob) of the optimized human adenovirus type 3 ciliary protein, the head structure of the ciliary protein of the optimized human adenovirus type 3 The amino acid sequence of the domain is shown in SEQ ID No:6.
[0046] S113. Artificially synthesizing the optimized gene sequence of the head domain (fiber knob) of the human adenovirus type 3 ciliary protein, amplifying and cloning it into an expression vector to obtain a recombinant expression vector...
Embodiment 1
[0100] Preparation of Ad3FK antigen
[0101] (1) The gene of the head domain (fiber knob) of the human adenovirus type 3 ciliary protein is codon-optimized, and the gene of the head domain (fiber knob) of the human adenovirus type 3 ciliary protein after optimization The amino acid sequence of the head domain of the ciliary protein of the optimized human adenovirus type 3 is shown in SEQ ID No:6.
[0102] (2) After amplifying the gene sequence of the head domain (fiber knob) of the human adenovirus type 3 ciliary protein optimized by artificial synthesis, an amplified product is obtained. The amplified product was double-digested with BamHI and HindIII and connected to the prokaryotic expression vector PQE30 (Qiagen Company) that had been double-digested with BamHI and HindIII to obtain a recombinant expression vector.
[0103] (3) Transform the recombinant expression vector into Escherichia coli BL21(DE3), and spread it on the r On the plate, cultivate for 12 hours, observe...
Embodiment 2
[0110] Preparation of Ad4FK, Ad7FK, Ad11FK, Ad14FK and Ad55FK
[0111] Head domain of ciliary protein of adenovirus type 4 (Ad4FK), head domain of ciliary protein of adenovirus type 7 (Ad7FK), head domain of ciliary protein of adenovirus type 11 (Ad11FK), type 14 The preparation method of the head domain (Ad14FK) of the ciliary protein of adenovirus and the head domain (Ad55FK) of the ciliary protein of type 55 adenovirus is roughly the same as the preparation method of the Ad3FK antigen in Example 1, and the difference is that the encoding The gene of each protein is different from Ad3FK antigen, and the amino acid constituting each protein is different from Ad3FK antigen. Specifically, the gene sequence encoding Ad4FK is shown in SEQ ID No: 7, and the amino acid sequence is shown in SEQ ID No: 8. The gene sequence encoding Ad7FK is shown in SEQ ID No:9, and the amino acid sequence is shown in SEQ ID No:10. The gene sequence encoding Ad11FK is shown in SEQ ID No:11, and t...
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