Effective screening of SARS-CoV-2 neutralizing antibodies in patient serum using lentivirus particles pseudotyped with SARS-CoV-2 spike glycoprotein

• Published in: Scientific reports 2020-11, Vol.10 (1), p.19076-19076, Article 19076
• Main Authors: Tandon, Ritesh, Mitra, Dipanwita, Sharma, Poonam, McCandless, Martin G., Stray, Stephen J., Bates, John T., Marshall, Gailen D.
• Format: Article
• Language: English
• Subjects: 631/326; 631/326/596; 692/308/2778; Animals; Antibodies, Neutralizing - blood; Antibodies, Neutralizing - immunology; Antibodies, Viral - blood; Antibodies, Viral - immunology; Betacoronavirus - immunology; Cell Line; Cell lines; COVID-19; Genetic Vectors - genetics; Glycoproteins; Humanities and Social Sciences; Humans; Immunoglobulin G; Immunoglobulin M; Lentivirus - genetics; Mammalian cells; multidisciplinary; SARS-CoV-2; Science; Science (multidisciplinary); Serologic Tests - methods; Severe acute respiratory syndrome coronavirus 2; Spike glycoprotein; Spike Glycoprotein, Coronavirus - genetics; Spike Glycoprotein, Coronavirus - immunology; Stomatitis; Transduction, Genetic
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-020-76135-w

Pseuodotyped particles have significant importance and use in virology as tools for studying the biology of highly pathogenic viruses in a lower biosafety environment. The biological, chemical, and serological studies of the recently emerged SARS-CoV-2 will be greatly aided by the development and optimization of a suitable pseudotyping system. Here, we pseudotyped the SARS-CoV-2 Spike glycoprotein (SPG) on a traditional retroviral (MMLV) as well as a third generation lentiviral (pLV) vector and tested the transduction efficiency in several mammalian cell lines expressing SARS-CoV-2 receptor hACE2. While MMLV pseudotyped the vesicular stomatitis virus G glycoprotein (VSV-G) efficiently, it could not pseudotype the full-length SPG. In contrast, pLV pseudotyped both glycoproteins efficiently; however, much higher titers of pLV-G particles were produced. Among all the tested mammalian cells, 293Ts expressing hACE2 were most efficiently transduced using the pLV-S system. The pLV-S particles were efficiently neutralized by diluted serum (>:640) from recently recovered COVID-19 patients who showed high SARS-CoV-2 specific IgM and IgG levels. In summary, pLV-S pseudotyped virus provides a valid screening tool for the presence of anti SARS-CoV-2 specific neutralizing antibodies in convalescent patient serum.