Development and optimized pairing of mouse monoclonal antibodies for detecting hemagglutinin in novel H7 subtype influenza viruses

Abstract

The H7 subtype avian influenza threatens public health with respect to poultry and humans. Thus, a specific and sensitive diagnostic test is essential for the management of H7 subtype influenza infections. In this study, five mouse monoclonal antibodies (mAbs) against hemagglutinin (HA) of influenza A/Anhui/1/2013 (H7N9) were produced and characterized by the Western blot, immunofluorescence, and hemagglutination inhibition assays. All five specific mAbs reacted with the HA protein of H7N9 but not with that of H1N1, H3N2, or H5N1. With the combination arrays of capture and detection antibodies, the matched pair mAbs (1C4-coated and 2D7-labeled) were selected and employed in a double-antibody sandwich ELISA (DAS-ELISA). Detection limits of the sandwich ELISA were 0.45 ng mL -1 for the HA protein derived from A/Anhui/1/2013 (H7N9); or 1 and 2 HA units/50 µL for A/Anhui/1/2013 (H7N9) and A/GD/17SF003/2016 (H7N9), respectively. These anti-HA mAbs against subtype H7 and the novel DAS-ELISA provide a valuable approach for specific detection of the H7 subtype influenza virus and quantification of its HA protein, especially for the novel epidemic H7N9.influenza A, H7 subtype, hemagglutinin, monoclonal antibodies, sandwich ELISA Tan, W. (2020). Development and optimized pairing of mouse monoclonal antibodies for detecting hemagglutinin in novel H7 subtype influenza viruses. Sci China Life Sci 63, 279-289. https://doi.

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

“…The advantages of using mAbs for detecting AIV include high specificity, sensitivity and unlimited provision of a standardized reagent. mAbs against the HA protein of AIV served as a useful tool to distinguish its subtypes in previous studies [22][23][24]. In the present paper, the cELISA immunoassay for H7 antibody detection was developed using an anti-H7-HA1 mAb targeting the epitope located in the depression under antigenic site E of H7-HA1 subunit [17].…”

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

“…6,8,14,18,19 A sandwich ELISA was developed with a detection limit of 0.45 ng/mL for the H7N9-HA protein derived from A/Anhui/1/2013 (H7N9), and 1 and 2 HAU/50 μL for live H7N9 AIVs. 20 An additional sandwich ELISA was developed with a lowest detection limit of 10 ng/mL for recombinant H7 protein and 0.5 −2 HAU/50 μL for live H7 AIVs, respectively. 21 T A B L E 1 Detection sensitivity of the H7N9 strip using clinical specimens Note: AC-ELISA and virus isolation methods are highly correlated (P < .001).…”

“…In recent years, AC‐ELISA has been applied widely to detect various AIVs including H5, H6, H7, H9N2, and H10N8 6,8,14,18,19 . A sandwich ELISA was developed with a detection limit of 0.45 ng/mL for the H7N9‐HA protein derived from A/Anhui/1/2013 (H7N9), and 1 and 2 HAU/50 μL for live H7N9 AIVs 20 . An additional sandwich ELISA was developed with a lowest detection limit of 10 ng/mL for recombinant H7 protein and 0.5 −2 HAU/50 μL for live H7 AIVs, respectively 21 .…”

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

“…The advantages of using mAbs for detecting AIV include high speci city, sensitivity and unlimited provision of a standardized reagent. mAbs against the HA protein of AIV served as a useful tool to distinguish its subtypes in previous studies [21][22][23]. In the present paper, the cELISA immunoassay for H7 antibody detection was developed using an anti-H7-HA1 mAb targeting the epitope located in the depression under antigenic site E of H7-HA1 subunit [17].…”