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Background
More than three million infections were attributed to Chikungunya virus (CHIKV) in the 2014–2016 outbreak in Mexico, Central and South America, with over 500 deaths directly or indirectly related to this viral disease. CHIKV outbreaks are recurrent and no vaccine nor approved therapeutics exist to prevent or treat CHIKV infection. Reliable and robust diagnostic methods are thus critical to control future CHIKV outbreaks. Direct CHIKV detection in serum samples via highly specific and high affinity anti-CHIKV antibodies has shown to be an early and effective clinical diagnosis.
Methods
Chikungunya virions isolated from serum of a patient in Veracruz, México, were purified and characterized via electron microscopy, SDS-PAGE and binding to diverse well-characterized anti-CHIKV monoclonal antibodies. UV-inactivated CHIKV particles were used as selector in a solid-phase panning coupled with ELISA-based screening and Next-Generation Sequencing to discover specific and high affinity anti-CHIKV antibodies from ALTHEA Gold Libraries™.
Results
The CHIKV isolate showed the typical morphology of the virus. Protein bands in the SDS-PAGE were consistent with the size of its capsid proteins. UV-inactivated CHIKV particles bound tightly the control antibodies. The lead antibodies here obtained showed high expression yield, monomeric content over 95% after a single-step Protein A purification, and importantly, a thermal stability above 75oC. Most of the antibodies recognized linear epitopes on E2, including the highest affinity antibody called C7. A sandwich ELISA implemented with C7 and a potent neutralizing antibody isolated elsewhere, also specific for E2 but recognizing a discontinuous epitope, showed a dynamic range of 0.2–40.0 µγ/mL of UV-inactivated CHIKV purified preparation. The number of CHIKV particles estimated based on the concentration of E2 in the extract suggested that the assay could detect clinically meaningful amounts of CHIKV in serum.
Conclusions
The newly discovered antibodies offer valuable tools for characterization of CHIKV isolates and development of robust diagnostic tools for CHIKV infection surveillance. Application of ALTHEA Gold Libraries™ in combination with viral particles other than CHIKV could expedite the discovery and development of antibodies for detection and control of emergent and quickly spreading viral outbreaks such as SARS-CoV-2 (COVID-19).
Keywords
Chikungunya virus, Venezuelan equine encephalitis Virus, Diagnostic, Human Antibodies, Next-Generation Sequencing, Phage display
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CURRENT STATUS: Under Review
The most recent version of this article is available here.
No community comments so far
Reviewers invited
Invitations sent on 10 Feb, 2021
Editor assigned
On 18 Nov, 2020
Submission checks complete
On 18 Nov, 2020
Editor invited
On 18 Nov, 2020
Version 1
Posted 19 Jun, 2020
No comments provided
Editorial decision: Major revision
On 13 Oct, 2020
Review #2 received
Received 05 Oct, 2020
Reviewer #2 agreed
On 14 Sep, 2020
Review #1 received
Received 11 Sep, 2020
Reviewer #1 agreed
On 09 Sep, 2020
Reviewers invited
Invitations sent on 06 Jul, 2020
Editor assigned
On 18 Jun, 2020
Submission checks complete
On 17 Jun, 2020
Editor invited
On 17 Jun, 2020
First submitted
On 13 Jun, 2020
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Subject Areas
Infectious Diseases
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This preprint is under consideration at BMC Infectious Diseases. A preprint is 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 article
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: Under Review
The most recent version of this article is available here.
No community comments so far
Reviewers invited
Invitations sent on 10 Feb, 2021
Editor assigned
On 18 Nov, 2020
Submission checks complete
On 18 Nov, 2020
Editor invited
On 18 Nov, 2020
Version 1
Posted 19 Jun, 2020
No comments provided
Editorial decision: Major revision
On 13 Oct, 2020
Review #2 received
Received 05 Oct, 2020
Reviewer #2 agreed
On 14 Sep, 2020
Review #1 received
Received 11 Sep, 2020
Reviewer #1 agreed
On 09 Sep, 2020
Reviewers invited
Invitations sent on 06 Jul, 2020
Editor assigned
On 18 Jun, 2020
Submission checks complete
On 17 Jun, 2020
Editor invited
On 17 Jun, 2020
First submitted
On 13 Jun, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Infectious Diseases
License:
This work is licensed under a CC BY 4.0 License. Read Full License
The most recent version of this article is available here.
Background
More than three million infections were attributed to Chikungunya virus (CHIKV) in the 2014–2016 outbreak in Mexico, Central and South America, with over 500 deaths directly or indirectly related to this viral disease. CHIKV outbreaks are recurrent and no vaccine nor approved therapeutics exist to prevent or treat CHIKV infection. Reliable and robust diagnostic methods are thus critical to control future CHIKV outbreaks. Direct CHIKV detection in serum samples via highly specific and high affinity anti-CHIKV antibodies has shown to be an early and effective clinical diagnosis.
Methods
Chikungunya virions isolated from serum of a patient in Veracruz, México, were purified and characterized via electron microscopy, SDS-PAGE and binding to diverse well-characterized anti-CHIKV monoclonal antibodies. UV-inactivated CHIKV particles were used as selector in a solid-phase panning coupled with ELISA-based screening and Next-Generation Sequencing to discover specific and high affinity anti-CHIKV antibodies from ALTHEA Gold Libraries™.
Results
The CHIKV isolate showed the typical morphology of the virus. Protein bands in the SDS-PAGE were consistent with the size of its capsid proteins. UV-inactivated CHIKV particles bound tightly the control antibodies. The lead antibodies here obtained showed high expression yield, monomeric content over 95% after a single-step Protein A purification, and importantly, a thermal stability above 75oC. Most of the antibodies recognized linear epitopes on E2, including the highest affinity antibody called C7. A sandwich ELISA implemented with C7 and a potent neutralizing antibody isolated elsewhere, also specific for E2 but recognizing a discontinuous epitope, showed a dynamic range of 0.2–40.0 µγ/mL of UV-inactivated CHIKV purified preparation. The number of CHIKV particles estimated based on the concentration of E2 in the extract suggested that the assay could detect clinically meaningful amounts of CHIKV in serum.
Conclusions
The newly discovered antibodies offer valuable tools for characterization of CHIKV isolates and development of robust diagnostic tools for CHIKV infection surveillance. Application of ALTHEA Gold Libraries™ in combination with viral particles other than CHIKV could expedite the discovery and development of antibodies for detection and control of emergent and quickly spreading viral outbreaks such as SARS-CoV-2 (COVID-19).
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
This is a list of supplementary files associated with this preprint. Click to download.
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