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Human anti-EV71 (enterovirus 71) neutralizing antibody EV71FabK7 and preparation method and application thereof

A virus and antibody technology, applied in the direction of antiviral agents, antiviral immunoglobulin, botany equipment and methods, etc., can solve the problems of restriction, blood-borne disease infection, time-consuming, etc.

Inactive Publication Date: 2012-10-10
INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] As an antibody component, immunoglobulin (Ig) mainly comes from the immune serum of donors (patients in the convalescence period). It takes a long time from obtaining positive serum to passing safety testing, and requires a lot of manpower and financial resources, which makes it Large-scale production is limited, and because the antibody is mainly derived from serum, it is prone to infection of blood-borne diseases

Method used

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  • Human anti-EV71 (enterovirus 71) neutralizing antibody EV71FabK7 and preparation method and application thereof
  • Human anti-EV71 (enterovirus 71) neutralizing antibody EV71FabK7 and preparation method and application thereof
  • Human anti-EV71 (enterovirus 71) neutralizing antibody EV71FabK7 and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Construction of Human Anti-EV71 Virus Antibody Library and Screening of Fab Antibody

[0033] 1.1 Materials and methods

[0034]1.1.1 Sources of viruses, cells and vectors: EV71 virus SHZH98 strain and FUYANG-0805 strain have been published in the literature (Yang F, Jin Q, He Y, Li L, Hou Y.2001. The complete genome of Enterovirus 71 China strain. Sci China C Life Sci and Wu Z, Yang F, Zhao R, Zhao L, Guo D, Jin Q.2009. Identification of small interfering RNAs which inhibit the replication of several Enterovirus 71 strains in China. J Virol Methods), by Provided by Institute of Pathogen Biology, Chinese Academy of Medical Sciences. The cells used for EV71 virus neutralization experiment in vitro were RD cells, which were purchased from ATCC. The bacterial strain is XL1-Blue (Stratagene, USA), and the vector pComb3H (provided by Scripps Research Institute, USA).

[0035] 1.1.2 Antigen preparation

[0036] Purification of EV71 virus: The culture supernatant...

Embodiment 2

[0109] Example 2 Application of Human Anti-EV71 Virus Neutralizing Antibody EV71FabK7

[0110] At present, there are no specific vaccines and drugs for the prevention and treatment of hand, foot and mouth disease. The immunoglobulins used clinically are all derived from the positive serum of human anti-EV71, which limits their large-scale preparation. Susceptible to infection with blood-borne diseases. The human source anti-EV71 virus genetically engineered antibody obtained by the invention is used to replace the blood source immunoglobulin, which provides a new approach for the treatment of hand, foot and mouth disease.

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Abstract

The invention provides human anti-EV71 (enterovirus 71) neutralizing antibody EV71FabK7 which is obtained by screening by means of phage display. Amino acid sequences of light chain and heavy chain variable areas are shown as SEQ ID No.1 and SEQ ID No.2 respectively. The antibody can specifically recognize EV71 virus particle antigens, can have evident enzyme-linked immune reaction with EV71 viruses, and has a function of neutralizing activity resistant to EV71 virus infection. Further, the antibody can be made into specific antibody medicine for preventing and treating the hand-foot-mouth disease and clinically used for preventing and treating the hand-foot-mouth disease caused by EV71 viruses.

Description

technical field [0001] The invention relates to genetic engineering antibody technology, in particular to a human-sourced anti-EV71 virus neutralizing antibody EV71FabK7, its preparation method and application. Background technique [0002] Hand foot mouth disease (Hand foot mouth disease, HFMD) is an infectious disease caused by enterovirus, mostly occurs in infants under 5 years old, can cause fever, rashes and ulcers on hands, feet, mouth and other parts, individual patients may Cause myocarditis, pulmonary edema, aseptic meningoencephalitis and other complications. There are more than 20 kinds of enteroviruses that cause HFMD, among which Cox Asckievirus A16 (Cox A16) and Enterovirus 71 (Enterovirus71.EV71) are the most common. In 1974, Schmidt et al. reported for the first time that EV71 was isolated from patients with neurological symptoms (1969-1973) that broke out in California, USA. Subsequently, many countries in the world successively reported the prevalence of E...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C12N15/13C12N15/63C12N1/21C12N1/19C12N5/10A61K39/42A61P31/14G01N33/569
Inventor 陈哲金奇
Owner INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI
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