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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

Active Publication Date: 2018-12-28
THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently no specific and effective treatment for human adenovirus type 3.

Method used

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  • Hybridoma cell capable of secreting anti-Ad3FK monoclonal antibody, monoclonal antibody and preparation method and application thereof
  • Hybridoma cell capable of secreting anti-Ad3FK monoclonal antibody, monoclonal antibody and preparation method and application thereof
  • Hybridoma cell capable of secreting anti-Ad3FK monoclonal antibody, monoclonal antibody and preparation method and application thereof

Examples

Experimental program
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preparation example Construction

[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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Abstract

The invention relates to a hybridoma cell capable of secreting an anti-Ad3FK monoclonal antibody, the monoclonal antibody and a preparation method and an application thereof. The hybridoma cell capable of secreting the anti-Ad3FK monoclonal antibody has a preservation number of CCTCC No: C201875. The anti-Ad3FK monoclonal antibody secreted by the hybridoma cell has the advantages of high specificity and stable performance, can specifically bind human adenovirus 3, can be widely used in preparation of a drug for treating relevant diseases infected with the human adenovirus 3, and can also be used in the preparation of a detection reagent or a detection device for detecting the human adenovirus 3.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell capable of secreting an anti-Ad3FK monoclonal antibody, a monoclonal antibody and a preparation method and application thereof. Background technique [0002] There are more than 50 serotypes and more than 69 genotypes of human adenovirus (human Adenovirus, HAdV) found so far in 7 groups. Adenovirus is one of the main pathogens causing viral pneumonia. Adenoviruses of group B, C, and E can all cause respiratory tract infections. Among them, human group B adenovirus types 3, 7, 14, 55, and E Adenovirus type 4 is an important pathogen that causes acute respiratory infection (ARI) and fatal pneumonia in infants and young children. It is highly contagious and often leads to outbreaks. [0003] The popular strains of human adenovirus in China are mainly type 3 adenovirus, especially in infants and young children, human type 3 adenovirus is the most important pathogen caus...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/08C12N15/13G01N33/577C12N15/06A61K39/395A61P31/20
CPCA61K2039/505A61P31/20C07K16/081C12N15/02G01N33/577
Inventor 田新贵周荣张玲
Owner THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)