Development of Monoclonal Antibodies Against SARS-CoV-2 Nucleocapsid Protein for COVID-19 Antigen Detection DOI

Maurine Mumo Mutua,

Bernard N. Kanoi, Steven Ger Nyanjom

и другие.

Research Square (Research Square), Год журнала: 2025, Номер unknown

Опубликована: Апрель 14, 2025

Abstract Background The coronavirus disease 2019 (COVID-19) pandemic underscored the global need for reliable diagnostic tools with quick turnaround time effective patient management and mitigation of virus spread. This study aimed to express severe acute respiratory syndrome 2 (SARS-CoV-2) nucleocapsid protein produce monoclonal antibodies (mAbs) against expressed protein. Methods Following successful expression purification His-tagged SARS-CoV-2 N using a wheat germ cell-free system (WGCFS), BALB/c mice were immunized, generated hybridomas screened mAb production. Indirect sandwich ELISA used screen reactivity antibody both our recombinant antigen commercial antigen. mAbs also assessed their performance RT-PCR confirmed positive samples varying cycle threshold (CT) values specificity intracellular fluid (ICF) other viruses. Results Our demonstrated high antigen, Beta Omicron variants. There was no significant difference in binding affinity (p = 0.12) 0.072) antigens. detected from clinical CT exhibited cross-reactivity Conclusion We successfully leveraging WGCFS resource-limited setting. had making it suitable candidate detection kit development. Beyond diagnostics, holds potential therapeutic applications as well use environmental surveillance platforms.

Язык: Английский

Development of Monoclonal Antibodies Against SARS-CoV-2 Nucleocapsid Protein for COVID-19 Antigen Detection DOI

Maurine Mumo Mutua,

Bernard N. Kanoi, Steven Ger Nyanjom

и другие.

Research Square (Research Square), Год журнала: 2025, Номер unknown

Опубликована: Апрель 14, 2025

Abstract Background The coronavirus disease 2019 (COVID-19) pandemic underscored the global need for reliable diagnostic tools with quick turnaround time effective patient management and mitigation of virus spread. This study aimed to express severe acute respiratory syndrome 2 (SARS-CoV-2) nucleocapsid protein produce monoclonal antibodies (mAbs) against expressed protein. Methods Following successful expression purification His-tagged SARS-CoV-2 N using a wheat germ cell-free system (WGCFS), BALB/c mice were immunized, generated hybridomas screened mAb production. Indirect sandwich ELISA used screen reactivity antibody both our recombinant antigen commercial antigen. mAbs also assessed their performance RT-PCR confirmed positive samples varying cycle threshold (CT) values specificity intracellular fluid (ICF) other viruses. Results Our demonstrated high antigen, Beta Omicron variants. There was no significant difference in binding affinity (p = 0.12) 0.072) antigens. detected from clinical CT exhibited cross-reactivity Conclusion We successfully leveraging WGCFS resource-limited setting. had making it suitable candidate detection kit development. Beyond diagnostics, holds potential therapeutic applications as well use environmental surveillance platforms.

Язык: Английский

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