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Humanized anti-Yersinia pestis antigen F1 antibody and application

An antibody and monoclonal antibody technology, applied in the direction of antibodies, applications, antibacterial drugs, etc., can solve the problems of unsustainable use and adverse reactions of antibiotics, and achieve the effect of high protection, high specificity and high affinity

Active Publication Date: 2020-09-22
中国人民解放军东部战区疾病预防控制中心
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  • Application Information

AI Technical Summary

Problems solved by technology

Vaccines must be inoculated within a certain period of time before exposure, and generally require multiple vaccinations to be effective. A large number of antibiotics can be used to treat plague patients and control the spread of the disease. Yersinia pestis has low resistance to antibiotics, but currently Yersinia pestis, which acquired multi-drug resistance by transferring plasmids, has been isolated in Madagascar, and antibiotics have certain adverse reactions and cannot be used continuously

Method used

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  • Humanized anti-Yersinia pestis antigen F1 antibody and application
  • Humanized anti-Yersinia pestis antigen F1 antibody and application
  • Humanized anti-Yersinia pestis antigen F1 antibody and application

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Embodiment 1

[0020] The preparation of embodiment 1hm YF202 antibody

[0021] After the detection of mouse monoclonal antibody in the early stage of the research group, we selected the YF202 hybridoma cell line to prepare the human-mouse chimeric antibody hm YF202.

[0022] 1) Amplification and verification of antibody variable region gene fragments:

[0023] The YF202 antibody hybridoma cells were cultured to the logarithmic growth phase, and the total RNA was extracted by Trizol-chloroform-isopropanol method; the dried total RNA was dissolved in 20 μL water, and the OD260 / OD280 was measured, and the value was 1.9. Take 14 μL of RNA for reverse transcription, use the mRNA in the total RNA as a template, and use OligodT 15 Single-stranded cDNA was obtained by reverse transcription and amplification.

[0024] Design 19 VH upstream primers and 17 Vκ upstream primers, 4 VH downstream primers and 3 Vκ downstream primers Primers:

[0025] Vκ5' upstream primer:

[0026] Vκ-1

[0027] 5’-GGG...

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Abstract

The invention discloses a humanized anti-Yersinia pestis antigen F1 antibody and application. A light chain variable region of the antibody has a nucleotide sequence SEQ ID NO: 1 and an amino acid sequence SEQ ID NO: 2, wherein a light chain hypervariable region sequence has SEQ ID NO: 3, SEQ ID NO:4 and SEQ ID NO:5; a heavy chain variable region has a nucleotide sequence SEQ ID NO: 6 and an aminoacid sequence SEQ ID NO: 7; and a heavy chain hypervariable region has SEQ ID NO: 8, SEQ ID NO: 9 and SEQ ID NO: 10. The antibody can be specifically combined with a Yersinia pestis antigen (F1).

Description

technical field [0001] The invention relates to the gene of anti-anthrax monoclonal antibody, the polypeptide encoded by the gene, the preparation technology of mouse monoclonal antibody and human-mouse chimeric antibody, and the application of the gene and polypeptide in the preparation of anthrax treatment and preventive medicine. Background technique [0002] Plague is a severe infectious disease caused by Yersinia pestis infection. The disease develops rapidly. If it is not treated in time, the mortality rate is extremely high. It has caused hundreds of millions of deaths in history. Human infection with plague is mainly due to contact with sick rodents or the bite of infected fleas, which can cause bubonic plague, which can develop into plague septicemia or secondary pneumonic plague, often causing human plague epidemics. In recent years, plague has been prevalent mainly in some areas of China, Africa, South America and India. Although the current incidence of plague i...

Claims

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Application Information

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IPC IPC(8): C07K16/12C12N15/13A61K39/40A61P31/04
CPCC07K16/1228A61P31/04C07K2317/24C07K2317/56C07K2317/565C07K2317/92A61K2039/505Y02A50/30
Inventor 朱进周婷婷林红杨展
Owner 中国人民解放军东部战区疾病预防控制中心
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