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Broad spectrum monoclonal antibody for identification of influenza virus hemagglutinin protein HA1 structural domains

A monoclonal antibody, hemagglutinin protein technology, applied in the direction of antiviral immunoglobulins, antibodies, antiviral agents, etc., can solve the problem that the neutralization activity is limited to a few strains of each subtype.

Active Publication Date: 2014-10-15
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 2008, Yoshida et al. reported a HA1 broad-spectrum neutralizing monoclonal antibody S139 / 1, which can neutralize influenza viruses of H1, H2, H3 and H13 subtypes, but the neutralizing activity of this monoclonal antibody is limited to each subtype. A few strains of the type (Yoshida R. et al., PLoS Pathog. 2009, 5: e1000350)

Method used

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  • Broad spectrum monoclonal antibody for identification of influenza virus hemagglutinin protein HA1 structural domains
  • Broad spectrum monoclonal antibody for identification of influenza virus hemagglutinin protein HA1 structural domains
  • Broad spectrum monoclonal antibody for identification of influenza virus hemagglutinin protein HA1 structural domains

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] Example 1: Preparation of anti-H1 subtype influenza virus HA monoclonal antibody (mAb)

[0088] (1) Preparation of virus:

[0089] Three seasonal H1N1 influenza virus strains A / New Calidonia / 20 / 1999 (H1N1), A / Xiamen / 3118 / 2006 (H1N1), A / Xiamen / N585 / 2009 (H1N1), a new influenza A H1N1 Virus strain A / California / 04 / 2009 (H1N1) was inoculated into MDCK cells, and after incubation at 37°C for 2 days, the supernatant was collected to obtain the amplified virus. Collect the live virus and inactivate it with 0.03% formalin at 4°C. The inactivated virus is detected by HA to determine the titer of the inactivated virus solution (Note: For the specific methods of HA titer determination and HI detection, refer to WHO operation Guide, A / California / 04 / 2009 was donated by the Department of Microbiology, University of Hong Kong, A / New Calidonia / 20 / 1999, A / Xiamen / 3118 / 2006, A / Xiamen / N585 / 2009 are all strains preserved in our laboratory).

[0090] (2) Experimental mice:

[0091] 6-week...

Embodiment 2

[0106] Example 2: The hemagglutination inhibition test method identifies the broad-spectrum reactivity of the H1 / H5 subtype hemagglutinin broad-spectrum monoclonal antibody

[0107] Select representative strains of seasonal H1N1 influenza virus, representative strains of new type A H1N1 influenza virus, representative strains of highly pathogenic H5N1 avian influenza virus and representative strains of seasonal H3N2 influenza virus isolated from different times, different regions, and representing different mutation types, The cross-reactivity of the monoclonal antibody ascites of the above 25 monoclonal antibody strains to different H subtype influenza viruses was identified by hemagglutination inhibition assay (HI). According to the reactivity of mAbs with various influenza virus strains, three broad-spectrum mAbs across H subtypes, 12H5, G10D10, and 8F11, which can simultaneously recognize H1 and H5 subtype influenza viruses, were identified (Table 1). Monoclonal antibody 1...

Embodiment 3

[0112] Example 3. Microwell cell neutralization assay method to identify broad-spectrum neutralization activity (Neutralization titer) of broad-spectrum monoclonal antibody across H1 / H5 subtype hemagglutinin

[0113] Neutralizing activity is an important indicator to evaluate whether a monoclonal antibody has therapeutic potential. The neutralization activity of the monoclonal antibodies 12H5, G10D10 and 8F11 against H1N1 and H5N1 subtype influenza virus representative strains was detected by microwell cell neutralization assay (Hulse-Post et al., PNAS.2005, 102:10682-7), and the results (Table 2) shows that the results of the neutralization test of monoclonal antibodies are basically consistent with the results of the hemagglutination inhibition test, and the three monoclonal antibodies (12H5, G10D10 and 8F11) all have broad-spectrum cross-neutralizing activities against H1N1 and H5N1 viruses.

[0114] Table 2 Neutralization test activity of broad-spectrum monoclonal antibody...

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Abstract

The invention relates to an antibody for identification of epitopes on influenza virus hemagglutinin protein HA1 structural domains, a cell strain generating the antibody and the use of the antibody and the cell strain. The antibody is can cross the HA subtype for specific binding of the hemagglutinin (HA) protein HA1 structural domains of H1 subtype (including seasonal H1N1 and new H1N1) and H5 subtype influenza virus. Therefore, the invention also relates vaccines or pharmaceutical compositions which comprise the antibody and are used for prevention and / or treatment of H1 subtype and H5 subtype influenza virus infection and / or diseases (such as the flu) induced by the H1 subtype and H5 subtype influenza virus.

Description

technical field [0001] The invention relates to the field of molecular virology, especially the field of vaccine prevention of influenza virus. Specifically, the present invention relates to an antibody that recognizes an epitope on the HA1 domain of the hemagglutinin protein of influenza virus, a cell line producing the antibody, and their use. The antibody of the present invention can specifically bind to the HA1 domain of the hemagglutinin (HA) protein of H1 subtype (including seasonal H1N1 and new A H1N1) and H5 subtype influenza virus across HA subtypes. Therefore, the present invention also relates to vaccines or pharmaceutical compositions useful for preventing and / or treating infection of H1 subtype and H5 subtype influenza virus and / or diseases (such as influenza) caused by said infection, which comprise the Antibody. Background technique [0002] Influenza, referred to as influenza, is a respiratory infectious disease caused by influenza virus. Its clinical chara...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C12N15/13C12N15/63C12N5/20A61K39/145A61K48/00G01N33/569A61P31/16A61K39/42C12P21/08G01N33/68G01N33/577C12R1/91
CPCA61K39/42C12N15/63C07K16/10A61K2039/505A61P31/16C07K16/1018C07K2317/33C07K2317/34C07K2317/76C07K2317/565G01N33/56983G01N2333/11G01N2469/10
Inventor 陈毅歆郑清炳李睿沈晨光柯少君罗文新陈鸿霖夏宁邵
Owner XIAMEN UNIV
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