A monoclonal antibody recognizes a highly conserved neutralizing epitope on hemagglutinin of H6N1 avian influenza virus

Abstract Background H7 subtype avian influenza has caused great concern in the global poultry industry and public health. The conventional serological subtype-specific diagnostics is implemented by hemagglutination inhibition (HI) assay despite lengthy operation time. In this study, an efficient, rapid and high-throughput competitive enzyme-linked immunosorbent assay (cELISA) was developed for detection of antibodies against H7 avian influenza virus (AIV) based on a novel monoclonal antibody specific to the hemagglutinin (HA) protein of H7 AIV. Results The reaction parameters including antigen coating concentration, monoclonal antibody concentration and serum dilution ratio were optimized for H7 antibody detection. The specificity of the cELISA was tested using antisera against H1 ~ H9, H11 ~ H14 AIVs and other avian viruses. The selected cut-off values of inhibition rates for chicken, duck and peacock sera were 30.11, 26.85 and 45.66% by receiver-operating characteristic (ROC) curve analysis, respectively. With HI test as the reference method, the minimum detection limits for chicken, duck and peacock positive serum reached 20, 21 and 2− 1 HI titer, respectively. Compared to HI test, the diagnostic accuracy reached 100, 98.6, and 99.3% for chicken, duck and peacock by testing a total of 400 clinical serum samples, respectively. Conclusions In summary, the cELISA assay developed in this study provided a reliable, specific, sensitive and species-independent serological technique for rapid detection of H7 antibody, which was applicable for large-scale serological surveillance and vaccination efficacy evaluation programs.

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Prediction of a common neutralizing epitope of H5N1 avian influenza virus by in silico molecular docking

Science Bulletin ◽

10.1007/s11434-008-0161-4 ◽

2008 ◽

Vol 53 (6) ◽

pp. 868-877 ◽

Cited By ~ 2

Author(s):

YuanQing Yan ◽

ShaoWei Li ◽

ChunYan Yang ◽

WenXin Luo ◽

MingQiao Wang ◽

...

Keyword(s):

Influenza Virus ◽

Molecular Docking ◽

Avian Influenza ◽

Avian Influenza Virus ◽

In Silico ◽

Neutralizing Epitope ◽

H5n1 Avian Influenza Virus ◽

H5n1 Avian Influenza ◽

In Silico Molecular Docking

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Monoclonal Antibody Preparation Against Nucleoprotein of Avian Influenza Virus Subtype H9N2

Iranian Red Crescent Medical Journal ◽

10.5812/ircmj.95906 ◽

2019 ◽

Vol 21 (10) ◽

Author(s):

Maryam Hashemi ◽

Rasool Madani ◽

Mahmoudreza Aghamaali ◽

Tara Emami ◽

Fariba Golchinfar

Keyword(s):

Monoclonal Antibody ◽

Influenza Virus ◽

Avian Influenza ◽

Avian Influenza Virus ◽

Antibody Preparation ◽

Avian Influenza Virus Subtype ◽

Virus Subtype

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Development and evaluation of a monoclonal antibody-based competitive ELISA for the detection of antibodies against H7 avian influenza virus

10.21203/rs.3.rs-36939/v2 ◽

2020 ◽

Author(s):

Yuan Li ◽

Hongliu Ye ◽

Meng Liu ◽

Suquan Song ◽

Jin Chen ◽

...

Keyword(s):

Monoclonal Antibody ◽

Influenza Virus ◽

Avian Influenza ◽

Avian Influenza Virus ◽

Large Scale ◽

Operation Time ◽

Reference Method ◽

Antibody Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Antibody Concentration

Abstract Background: H7 subtype avian influenza have caused great concern in the global poultry industry and public health. The conventional serological subtype-specific diagnostics is implemented by hemagglutination inhibition (HI) assay despite lengthy operation time. In this study, an efficient, rapid and high-throughput competitive enzyme-linked immunosorbent assay (cELISA) was developed for detection of antibodies against H7 avian influenza virus (AIV) based on a novel monoclonal antibody specific to the hemagglutinin (HA) protein of H7 AIV. Results: The reaction parameters including antigen coating concentration, monoclonal antibody concentration and serum dilution ratio were optimized for H7 antibody detection. The specificity of the cELISA was tested using antisera against H1~H9, H11~H14 AIVs and other avian viruses. The selected cut-off values of inhibition rates for chicken, duck and peacock sera were 30.11%, 26.85% and 45.66% by receiver-operating characteristic (ROC) curve analysis, respectively. With HI test as the reference method, the minimum detection limits for chicken, duck and peacock positive serum reached 2 0 , 2 1 and 2 -1 HI titer, respectively. Compared to HI test, the diagnostic accuracy reached 100%, 98.6%, and 99.3% for chicken, duck and peacock by testing a total of 400 clinical serum samples, respectively. Conclusions: In summary, the cELISA assay developed in this study provided a reliable, specific, sensitive and species-independent serological technique for rapid detection of H7 antibody, which was applicable for large-scale serological surveillance and vaccination efficacy evaluation programs.

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Characterization of mAb6-9-1 monoclonal antibody against hemagglutinin of avian influenza virus H5N1 and its engineered derivative, single-chain variable fragment antibody

Acta Biochimica Polonica ◽

10.18388/abp.2016_1292 ◽

2016 ◽

Vol 64 (1) ◽

Cited By ~ 2

Author(s):

Róza Sawicka ◽

Paweł Siedlecki ◽

Barbara Kalenik ◽

Jan P Radomski ◽

Violetta Sączyńska ◽

...

Keyword(s):

Monoclonal Antibody ◽

Influenza Virus ◽

Avian Influenza ◽

Avian Influenza Virus ◽

Structural Model ◽

Expression System ◽

Antigenic Specificity ◽

Hybridoma Cells ◽

Single Chain Variable Fragment ◽

Single Chain

Hemagglutinin (HA), as a major surface antigen of influenza virus, is widely used as a target for production of neutralizing antibodies. Monoclonal antibody, mAb6-9-1, directed against HA of highly pathogenic avian influenza virus A/swan/Poland/305-135V08/2006(H5N1) was purified from mouse hybridoma cells culture and characterized. The antigenic specificity of mAb6-9-1 was verified by testing its cross-reactivity with several variants of HA. The mimotopes recognized by mAb6-9-1 were selected from two types of phage display libraries. The comparative structural model of the HA variant used for antibody generation was developed to further facilitated epitope mapping. Based on the sequences of the affinity-selected polypeptides and the structural model of HA the epitope has been located to the region near the receptor binding site (RBS). Such localization of the epitope recognized by mAb6-9-1 is in concordance with its moderate hemagglutination inhibition activity and its antigenic specificity. Additionally, total RNA from hybridoma cells secreting mAb6-9-1 was used for obtaining two variants of cDNA encoding recombinant single-chain variable fragment (scFv) antibody. To ensure high production level and solubility in bacterial expression system, the scFv fragments were produced as chimeric proteins in fusion with thioredoxin or displayed on a phage surface after cloning into the phagemid vector. Specificity and affinity of the recombinant soluble and phage-bound scFv were assayed by suitable variants of ELISA test. The observed slight differences in specificity are discussed.

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Complementary monoclonal antibody-based dot ELISA for universal detection of H5 avian influenza virus

BMC Microbiology ◽

10.1186/1471-2180-10-330 ◽

2010 ◽

Vol 10 (1) ◽

pp. 330 ◽

Cited By ~ 18

Author(s):

Fang He ◽

Retno D Soejoedono ◽

Sri Murtini ◽

Michael Goutama ◽

Jimmy Kwang

Keyword(s):

Monoclonal Antibody ◽

Influenza Virus ◽

Avian Influenza ◽

Avian Influenza Virus ◽

Universal Detection ◽

Dot Elisa

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Development of a monoclonal antibody‐based antigen capture enzyme‐linked immunosorbent assay for detection of H7N9 subtype avian influenza virus

Journal of Medical Virology ◽

10.1002/jmv.26292 ◽

2020 ◽

Author(s):

Fan Yang ◽

Yixin Xiao ◽

Fumin Liu ◽

Hangping Yao ◽

Nanping Wu ◽

...

Keyword(s):

Monoclonal Antibody ◽

Influenza Virus ◽

Avian Influenza ◽

Avian Influenza Virus ◽

Immunosorbent Assay ◽

Enzyme Linked Immunosorbent Assay ◽

Antigen Capture ◽

H7n9 Subtype

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Development and evaluation of a monoclonal antibody-based competitive ELISA for the detection of antibodies against H7 avian influenza virus

10.21203/rs.3.rs-36939/v1 ◽

2020 ◽

Author(s):

Yuan Li ◽

Hongliu Ye ◽

Meng Liu ◽

Suquan Song ◽

Jin Chen ◽

...

Keyword(s):

Monoclonal Antibody ◽

Influenza Virus ◽

Avian Influenza ◽

Avian Influenza Virus ◽

Large Scale ◽

Operation Time ◽

Reference Method ◽

Enzyme Linked Immunosorbent Assay ◽

Antibody Concentration ◽

Roc Curve Analysis

Abstract Background Since 2013, avian influenza outbreaks in poultry especially the H7 subtype have been causing a major concern for poultry industry and public health in China. The conventional serological subtype-specific diagnostics is implemented by hemagglutination inhibition (HI) assay despite lengthy operation time. An efficient, rapid and high-throughput competitive enzyme-linked immunosorbent assay (cELISA) was developed for detection of antibodies against H7 avian influenza virus (AIV) based on a novel monoclonal antibody specific to the hemagglutinin (HA) protein of H7 AIV. Results The reaction parameters of antigen coating concentration, monoclonal antibody concentration and serum dilution ratio were optimized for detection of H7 antibodies in chicken, ducks and peacocks sera. The specificity of the cELISA was tested using antisera against H1 ~ H9, H11 ~ H14 AIVs and other avian viruses. The selected cut-off values of inhibition rates for chicken, duck and peacock sera were 30.11%, 26.85% and 45.66% by receiver-operating characteristic (ROC) curve analysis, respectively. With the HI test as a reference method, the minimum detection limits for chicken, duck and peacock positive serum reached 20, 21 and 2− 1 HI titer, respectively. Compared to the HI test, the diagnostic accuracy reached 100%, 98.6%, and 99.3% for chicken, duck and peacock by testing a total of 400 clinical serum samples, respectively. Conclusions In summary, the cELISA immunoassay for rapid detection of H7 antibody in sera from different avian species was highly specific, extremely sensitive and might be exploited for the large-scale serological diagnosis of H7 AIVs.

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