See the published version of this preprint at Virology Journal.
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Christian Grund
Friedrich Loeffler Institute Federal Research Institute for Animal Health Institute of Bacterial Infections and Zoonoses: Friedrich-Loeffler-Institut Bundesforschungsinstitut fur Tiergesundheit Institut fur bakterielle Infektionen und Zoonosen
Corresponding Author
License:
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Background:
Newcastle disease is a devastating disease in poultry caused by Newcastle disease virus (NDV), a paramyxovirus endemic in many regions of the world despite intensive vaccination. Phylogenetic analysis reveal ongoing evolution of the predominant circulating genotype 2.VII, and the relevance of potential antigenic drift is under discussion. To investigate variation within neutralization-sensitive epitopes within the protein responsible for receptor binding, i.e. the Heamagglutinin-Neuraminidase (HN) spike protein, we were interested to established genotype-specific monoclonal antibodies (MAbs).
Methods:
An HN-enriched fraction of a gradient-purified NDV genotype 2.VII was prepared and successfully employed to induced antibodies in BalbC mice that recognize conformationally intact sites reactive by haemagglutination inhibition (HI). For subsequent screening of mouse hybridoma cultures, an NDV-ELISA was established that utilize Concanavalin A (ConA-ELISA) coupled Glycoproteins that was proven to present conformation-dependent epitopes.
Results:
Six out of nine selected MAbs were able to block receptor binding as demonstrated by HI-activity. One MAb recognized an epitope only present in the homologue virus while four other MAbs showed weak reactivity to selected other genotypes. On the other hand, one broadly cross-reacting MAb reacted with all genotypes tested and resembled the reactivity profile of genotype specific polyclonal antibody preparations that point to minor antigenic differences between tested NDV genotpyes.
Conclusions:
These results point to the concurrent presence of variable and conserved epitopes within the HN-molecule of NDV. The described protocol should help to generate MAbs to a variety of NDV strains and enable in depth analysis of the antigenic profiles of different genotypes.
Keywords
monoclonal antibody, Newcastle disease virus, genotype 2.VII, antigenicity, Haemagglutinin-Neuraminidase protein, conformational epitopes
Figure 1
Figure 2
Figure 3
Figure 4
This is a list of supplementary files associated with this preprint. Click to download.
- sfigure1ppt.pptx
Supplementary Figure 1: Antigenic profiling of different NDV genotypes Sera against five different genotypes were tested by HI against nine different antigens representing eight different NDV genotpyes. Boxplots represent results of three independent tests with three replicates each. Significant differences (p<0.05) to homologues serum, marked by red boxes, are indicated (*).
- Supp1.png
Supplemental Table 1: Antigenic relation of NDV genotypes
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CURRENT STATUS: Published
View the published article at Virology Journal.
Version 1
Posted 14 Oct, 2020
No community comments so far
Review #1 received
Received 01 Jan, 2021
Editorial decision: Minor revision
On 01 Jan, 2021
Review #2 received
Received 30 Dec, 2020
Review #3 received
Received 30 Dec, 2020
Reviewer #3 agreed
On 15 Dec, 2020
Reviewer #2 agreed
On 14 Dec, 2020
Reviewers invited
Invitations sent on 13 Dec, 2020
Reviewer #1 agreed
On 13 Dec, 2020
Editor invited
On 07 Oct, 2020
Editor assigned
On 06 Oct, 2020
Submission checks complete
On 06 Oct, 2020
First submitted
On 05 Oct, 2020
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Subject Areas
Virology
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See the published version of this preprint at Virology Journal.
This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research
Christian Grund
Friedrich Loeffler Institute Federal Research Institute for Animal Health Institute of Bacterial Infections and Zoonoses: Friedrich-Loeffler-Institut Bundesforschungsinstitut fur Tiergesundheit Institut fur bakterielle Infektionen und Zoonosen
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
View the published article at Virology Journal.
Version 1
Posted 14 Oct, 2020
No community comments so far
Review #1 received
Received 01 Jan, 2021
Editorial decision: Minor revision
On 01 Jan, 2021
Review #2 received
Received 30 Dec, 2020
Review #3 received
Received 30 Dec, 2020
Reviewer #3 agreed
On 15 Dec, 2020
Reviewer #2 agreed
On 14 Dec, 2020
Reviewers invited
Invitations sent on 13 Dec, 2020
Reviewer #1 agreed
On 13 Dec, 2020
Editor invited
On 07 Oct, 2020
Editor assigned
On 06 Oct, 2020
Submission checks complete
On 06 Oct, 2020
First submitted
On 05 Oct, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Virology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at Virology Journal.
Background:
Newcastle disease is a devastating disease in poultry caused by Newcastle disease virus (NDV), a paramyxovirus endemic in many regions of the world despite intensive vaccination. Phylogenetic analysis reveal ongoing evolution of the predominant circulating genotype 2.VII, and the relevance of potential antigenic drift is under discussion. To investigate variation within neutralization-sensitive epitopes within the protein responsible for receptor binding, i.e. the Heamagglutinin-Neuraminidase (HN) spike protein, we were interested to established genotype-specific monoclonal antibodies (MAbs).
Methods:
An HN-enriched fraction of a gradient-purified NDV genotype 2.VII was prepared and successfully employed to induced antibodies in BalbC mice that recognize conformationally intact sites reactive by haemagglutination inhibition (HI). For subsequent screening of mouse hybridoma cultures, an NDV-ELISA was established that utilize Concanavalin A (ConA-ELISA) coupled Glycoproteins that was proven to present conformation-dependent epitopes.
Results:
Six out of nine selected MAbs were able to block receptor binding as demonstrated by HI-activity. One MAb recognized an epitope only present in the homologue virus while four other MAbs showed weak reactivity to selected other genotypes. On the other hand, one broadly cross-reacting MAb reacted with all genotypes tested and resembled the reactivity profile of genotype specific polyclonal antibody preparations that point to minor antigenic differences between tested NDV genotpyes.
Conclusions:
These results point to the concurrent presence of variable and conserved epitopes within the HN-molecule of NDV. The described protocol should help to generate MAbs to a variety of NDV strains and enable in depth analysis of the antigenic profiles of different genotypes.
Figure 1
Figure 2
Figure 3
Figure 4
This is a list of supplementary files associated with this preprint. Click to download.
- sfigure1ppt.pptx
Supplementary Figure 1: Antigenic profiling of different NDV genotypes Sera against five different genotypes were tested by HI against nine different antigens representing eight different NDV genotpyes. Boxplots represent results of three independent tests with three replicates each. Significant differences (p<0.05) to homologues serum, marked by red boxes, are indicated (*).
- Supp1.png
Supplemental Table 1: Antigenic relation of NDV genotypes
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