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2014•China Animal Husbandry & Veterinary MedicineRequires access

Preparation of Monoclonal Antibody against Swine Influenza Virus HA by Nucleic Acid Immunization

Yu Li

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Abstract

For the preparation of monoclonal antibody(McAb)against HA protein of swine influenza virus,DNA plasmid bearing HAgene of A/Swine/Guangdong/2004(H1N1)was used to immunize BALB/c mice followed by generation of a McAb using the hybridoma technique.The generated nine McAbs were identified by ELISA.Functional analysis demonstrated the McAb 4D5 neutralized the virus.Furthermore,the McAb 4D5didn't have cross-reactivity with H3N2 or H5N1influenza viruses,showing highly specific to H1N1.Immunofluorescence assay showed that McAb 4D5 was able to detect infection of cells with H1N1.Over all,McAb 4D5 proved specific and highly sensitive for A/Swine/Guangdong/2004(H1N1)HA and could potentially serve as immunodiagnostic tools for the rapid detection of this specific strain of influenza virus and laid the foundation for the study of single chain antibody fragment.

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What this paper is about

For the preparation of monoclonal antibody(McAb)against HA protein of swine influenza virus,DNA plasmid bearing HAgene of A/Swine/Guangdong/2004(H1N1)was used to immunize BALB/c mice followed by generation of a McAb using the hybridoma technique.The generated nine McAbs were identified by ELISA.Functional analysis demonstrated the McAb 4D5 neutralized the virus.Furthermore,the McAb 4D5didn't have cross-reactivity with H3N2 or H5N1influenza viruses,showing highly specific to H1N1.Immunofluorescence assay showed that McAb 4D5 was able to detect infection of cells with H1N1.Over all,McAb 4D5 proved specific and highly sensitive for A/Swine/Guangdong/2004(H1N1)HA and could potentially serve as immunodiagnostic tools for the rapid detection of this specific strain of influenza virus and laid the foundation for the study of single chain antibody fragment.

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Available abstract

For the preparation of monoclonal antibody(McAb)against HA protein of swine influenza virus,DNA plasmid bearing HAgene of A/Swine/Guangdong/2004(H1N1)was used to immunize BALB/c mice followed by generation of a McAb using the hybridoma technique.The generated nine McAbs were identified by ELISA.Functional analysis demonstrated the McAb 4D5 neutralized the virus.Furthermore,the McAb 4D5didn't have cross-reactivity with H3N2 or H5N1influenza viruses,showing highly specific to H1N1.Immunofluorescence assay showed that McAb 4D5 was able to detect infection of cells with H1N1.Over all,McAb 4D5 proved specific and highly sensitive for A/Swine/Guangdong/2004(H1N1)HA and could potentially serve as immunodiagnostic tools for the rapid detection of this specific strain of influenza virus and laid the foundation for the study of single chain antibody fragment.

Key concepts: Monoclonal antibody, Virology, Virus, Immunofluorescence, Antibody, Immunization, Nucleic acid, Molecular biology

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