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Rapid quantitative screening assay for SARS-CoV-2 neutralizing antibodies using HiBiT-tagged virus-like particles

By Kei Miyakawa, Sundararaj Stanleyraj Jeremiah, Norihisa Ohtake, Satoko Matsunaga, Yutaro Yamaoka, Mayuko Nishi, Takeshi Morita, Ryo Saji, Mototsugu Nishii, Hirokazu Kimura, Hideki Hasegawa, Ichiro Takeuchi, Akihide Ryo

Posted 22 Jul 2020
medRxiv DOI: 10.1101/2020.07.20.20158410

SARS-CoV-2 neutralizing antibodies confer protective immunity against reinfection. We have developed a rapid test for screening SARS-CoV-2 neutralization antibodies using genome-free virus-like particles incorporated with a small luciferase peptide, HiBiT. Their entry into LgBiT-expressing target cells reconstitutes NanoLuc luciferase readily detected by a luminometer. This newly developed HiBiT-tagged Virus-like particle-based Neutralization Test (hiVNT) can readily quantify SARS-CoV-2 neutralizing antibodies within three hours with a high-throughput in a low biosafety setting. Moreover, the neutralizing activity obtained from hiVNT was highly consistent with that measured by the conventional neutralization test using authentic SARS-CoV-2. Furthermore, antibody responses to both viral spike and nucleocapsid proteins correlated with the neutralization activity assessed by hiVNT. Our newly-developed hiVNT could be instrumental to survey individuals for the presence of functional neutralizing antibody against SARS-CoV-2.

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