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Human source anti-rabies virus Fab antibody, cross linking method thereof with nanometer particles and application thereof

A nanoparticle and rabies virus technology, applied in the field of biomedicine, can solve problems such as patents or research reports on nano-antibody that have not yet been seen

Inactive Publication Date: 2013-04-10
中国人民解放军南京军区军事医学研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The applicant has searched domestic and foreign literature, but there is no report on the invention patent or research of nano-antibody in the field of rabies post-exposure prophylaxis.

Method used

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  • Human source anti-rabies virus Fab antibody, cross linking method thereof with nanometer particles and application thereof
  • Human source anti-rabies virus Fab antibody, cross linking method thereof with nanometer particles and application thereof
  • Human source anti-rabies virus Fab antibody, cross linking method thereof with nanometer particles and application thereof

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Embodiment 1

[0027] 1. Construction of fully human anti-rabies virus Fab phage antibody library

[0028] 1. Amplification of human Fab gene

[0029] Total RNA was extracted from peripheral blood lymphocytes of 45 healthy volunteers immunized with rabies vaccine, reverse-transcribed into cDNA, and specific primers of human antibodies were used (refer to Carlos F.Barbas III, Dennis R.Burton, Jamie K.Scott, Gregg J .Silverman.Phage display: A laboratory manual.1rd ed.New York: Gold spring harbor laboratory press, 2004) amplified V H , V κ , V λ gene, while amplifying C with the plasmid pComb3XTT H 1. C κ , using pComb3Xλ as a template to amplify C λ The Fab gene fragment was prepared through three overlapping PCRs; the above PCR products were recovered by agarose gel electrophoresis respectively, and after purification by the gel recovery kit, about 1600bp Fab fragments were obtained, see figure 1 .

[0030] 2. Establishment of anti-rabies virus phage antibody library

[0031] The Fab...

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Abstract

The invention relates to a human source anti-rabies virus Fab antibody, a cross linking method thereof with nanometer particles and application thereof. A VH chain of the antibody has an amino acid sequence shown by SEQ ID NO. 1, and in addition, a VL chain of the antibody has an amino acid sequence shown by SEQ ID NO. 3. The Fab antibody and a nanometer antibody Fab-CPNPs thereof of the anti-rabies viruses prepared by the invention attack Kunming mice through rabies virus standard conteracting toxic strains CVS-24, and at the same time, vaccine is given to be associated with Fab antibodies or nanometer Fab-CPNPs antibodies with different dosages for carrying out prevention experiments on the mice after the exposure. The results show that only the vaccine prevention is given, the sufficient protection can not be provided, and the Fab and the Fab-CPNPs with different dosages have a certain protection effect on the mice after the rabies exposure. The prepared nanometer antibody has potential application prospects in the field of treatment after the rabies exposure.

Description

1. Technical field [0001] The invention relates to the field of biomedicine, in particular to the preparation and application of a human-sourced anti-rabies virus Fab antibody cross-linked by nanoparticles. 2. Background technology [0002] Rabies is a zoonosis caused by Rabies virus. Mortality rate is as high as 100% in humans and animals. About 55,000 people die from rabies every year in the world, mainly in developing countries in Asia, Africa and Latin America. Human patients are mostly infected by the bites of sick animals, and have central nervous system infection diseases mainly characterized by hydrophobia, photophobia, dysphagia, and mania. The disease is widespread and has many hosts. Almost all warm-blooded animals can be infected. It takes various mammals as the main host. Infectious diseases are also one of the diseases that humans have not been able to effectively control, so it is very important to develop drugs for the prevention and treatment of rabies. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/10A61K39/44A61P31/14
Inventor 朱进刘新建林红冯振卿管晓虹
Owner 中国人民解放军南京军区军事医学研究所