H5 subtype bird-flue virus H5N1 hemagglutinin monoclonal antibody and nucleic acid sequence and preparation method thereof

Abstract

The invention discloses a nucleic acid sequence of a heavy-chain variable region and a nucleic acid sequence of a light-chain variable region of an H5 subtype bird-flue virus H5N1 hemagglutinin monoclonal antibody. The invention also discloses a nucleic acid sequence of a monoclonal antibody, which includes a nucleic acid sequence encoding the heavy-chain variable region and a nucleic acid sequence encoding the light-chain variable region. The invention also discloses a preparation method of the monoclonal antibody: H5 subtype bird-flue virus H5N1 hemagglutinin immune BALB / c mice are used for preparing a set of mouse anti-H5 hemagglutinin specificity monoclonal antibodies. Hybridoma plants 3F16A4 of high-titre neutralized activity are sieved out by an in-vitro neutralized activity test. The sequences of the heavy-chain variable region and the light-chain variable region of the monoclonal antibody are measured. The sequence encoding the variable region has a high potential value on the aspects of building the chimeric antibody, the humanization antibody, and the like which have treating functions on highly-pathogenic H5 subtype bird-flue virus H5N1.

Description

technical field [0001] The invention relates to a monoclonal antibody, in particular to a highly neutralizing active anti-H5 subtype avian influenza virus (H5N1) hemagglutinin-specific monoclonal antibody (3F16A4). The present invention also relates to a variable region gene of the above antibody. The present invention also relates to a preparation method of the above antibody. Background technique [0002] Highly pathogenic avian influenza poses a huge threat to public health. Since the first discovery of the highly pathogenic H5 subtype avian influenza virus H5N1 infecting humans in Hong Kong in 1997, an outbreak of the H5 subtype highly pathogenic avian influenza virus infecting humans occurred again in early 2003 in Hong Kong. Since 2004, H5N1 avian influenza virus infection incidents and highly pathogenic avian influenza virus avian infection outbreaks have occurred continuously in many countries. Capabilities are gradually increasing. Therefore, the World Health Or...

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Problems solved by technology

However, there is a 3F16A4 monoclonal antibody that has always maintained a high neutralizing titer against the highly pathogenic H5 subtype avian influenza virus H5N1 isolated from humans from 1997 to 2006, indicating that although the mutation frequency of the H5N1 virus is very high, there are many viruses. Among the neutralizing antigenic sites, there are still relatively conserved sites that have withstood enormous mutation pressure. This antigenic site may be a highly conserved neutralizing site, which is an important target for effective prevention and control of viruses. point

[0004] Chinese invention patent specification CN1814623A (disclosure date: August 9, 2006) discloses a patent application titled "a monoclonal antibody to the hemagglutinin protein of H5 subtype avian influenza virus, its preparation method and its use" , which discloses a monoclonal antibody 2F2 against H5 subtype avian influenza virus hemagglutinin protein and the sequences of its heavy chain variable region and light chain variable region, as well as the hybridoma cell CCTCC-C200424 producing the antibody , the inventor of the application published in November 2005 the monoclonal antibody 2F2 isolated from 1987 to 2005 in the article "Preparation and Preliminary Application of Highly Pathogenic H5 Subtype Avian Influenza Virus Hemagglutinin Monoclonal Antibody" The hemagglutination inhibition test of the H5 subtype avian influenza virus strains showed that the blood inhibitory titers for many viruses were higher than 1:6400, but for the virus strains Sck / ST / 4610 / 03 and Wdk / JK / The blood inhibitory titers of 1653 / 05 are low, 1:200 and less than 1:100 respectively, which shows that the antigenic site recognized by this antibody is not a conserved site and has mutated

Moreover, the H5 subtype viruses recognized by the monoclonal antibody are all isolated from poultry, and the H5N1 virus that can infect humans is unknown

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