Rigid monoclonal antibodies improve detection of SARS-CoV-2 nucleocapsid protein

Monoclonal antibodies (mAbs) are the basis of treatments and diagnostics for pathogens and other biological phenomena. We conducted a structural characterization of mAbs against the N-terminal domain of nucleocapsid protein (NP ) from SARS-CoV-2 using small angle X-ray scattering (SAXS). Our solutio...

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Bibliographic Details
Published in:bioRxiv 2021-01
Main Authors: Hodge, Curtis D, Rosenberg, Daniel J, Wilamowski, Mateusz, Joachimiak, Andrzej, Hura, Greg L, Hammel, Michal
Format: Article
Language:English
Subjects:
Antigens
Biophysics
Epidemiology
Epitopes
Monoclonal antibodies
Nucleocapsids
Pathogens
Polymerization
Severe acute respiratory syndrome coronavirus 2
X-ray scattering
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Summary:Monoclonal antibodies (mAbs) are the basis of treatments and diagnostics for pathogens and other biological phenomena. We conducted a structural characterization of mAbs against the N-terminal domain of nucleocapsid protein (NP ) from SARS-CoV-2 using small angle X-ray scattering (SAXS). Our solution-based results distinguished the mAbs' flexibility and how this flexibility impacts the assembly of multiple mAbs on an antigen. By pairing two mAbs that bind different epitopes on the NP , we show that flexible mAbs form a closed sandwich-like complex. With rigid mAbs, a juxtaposition of the Fabs is prevented, enforcing a linear arrangement of the mAb pair, which facilitates further mAb polymerization. In a modified sandwich ELISA, we show the rigid mAb-pairings with linear polymerization led to increased NP detection sensitivity. These enhancements can expedite the development of more sensitive and selective antigen-detecting point-of-care lateral flow devices (LFA), key for early diagnosis and epidemiological studies of SARS-CoV-2 and other pathogens.
ISSN:2692-8205
DOI:10.1101/2021.01.13.426597