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 (NPNTD) 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 NPNTD, 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 NPNTD 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.
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