Abstract
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 NTD ) from SARS-CoV-2 using small-angle X-ray scattering and transmission electron microscopy. Our solution-based results distinguished the mAbs’ flexibility and how this flexibility affects the assembly of multiple mAbs on an antigen. By pairing two mAbs that bind different epitopes on the NP NTD , we show that flexible mAbs form a closed sandwich-like complex. With rigid mAbs, a juxtaposition of the antigen-binding fragments is prevented, enforcing a linear arrangement of the mAb pair, which facilitates further mAb polymerization. In a modified sandwich enzyme-linked immunosorbent assay, we show that rigid mAb-pairings with linear polymerization led to increased NP NTD detection sensitivity. These enhancements can expedite the development of more sensitive and selective antigen-detecting point-of-care lateral flow devices, which are critical for early diagnosis and epidemiological studies of SARS-CoV-2 and other pathogens.
Keywords: SARS-cov-2; flexibility; mAbs polymerization; nucleocapsid; small-angle x-ray scattering.
【저자키워드】 SARS-CoV-2, nucleocapsid, flexibility, mAbs polymerization, small-angle x-ray scattering., 【초록키워드】 Treatment, antibody, monoclonal antibodies, diagnostics, nucleocapsid protein, Antigen, lateral flow, early diagnosis, point-of-care, enzyme-linked immunosorbent assay, Epitopes, pathogen, nucleocapsid, Pathogens, NTD, epitope, transmission electron microscopy, Critical, mAbs, assembly, mAb, flexibility, N-terminal domain, Detection sensitivity, epidemiological studies, complex, Epidemiological study, selective, Affect, enzyme-linked immunosorbent, juxtaposition, flexible, conducted, linear, facilitate, prevented, X-ray scattering, biological phenomena, scattering, 【제목키워드】 monoclonal antibody, Protein, SARS-CoV-2 nucleocapsid, IMPROVE,