Large scale production and characterization of SARS‐CoV‐2 whole antigen for serological test development

Abstract

On December 31 st 2019, China notified the World Health Organization (WHO) of a novel coronavirus outbreak which was first observed a few weeks earlier in a patient presenting with severe respiratory disease in Wuhan. 1 In January 2020, the virus was identified as a novel coronavirus with more than 70% similarity with the severe acute respiratory syndrome coronavirus (SARS-CoV), and it was officially named by the WHO as 2019 novel coronavirus (nCoV) or SARS-CoV-2. 1 Currently, SARS-CoV-2 has attributed to a worldwide pandemic resulting in over 40 million infections and 1 million deaths, with numbers constantly growing. 2 Coronaviruses are enveloped, non-segmented, positive-sense single-stranded RNA viruses with a diameter of 60-140 nm, including the spike, with genome sizes ranging from 26 to 32 kilobases (the largest known viral RNA genome). 3,4 Coronaviruses have four

Effect of Inactivation Methods on SARS-CoV-2 Virion Protein and Structure

“…Jureka et al demonstrate that 56 • C inactivation requires more than 30 min exposure, while 100 • C incubation is effective within 5 min. Our findings at 65 • C are in line with prior published reports for SARS-CoV-1 and 2 [2,9,14]. Despite differences in susceptibility or extent, we have identified and compared conditions of UV and heat inactivation that effectively inactivated SARS-CoV-2.…”

“…Studies like Jureka et al demonstrated excellent inactivation with Trizol, formalin and beta-propiolactone (BPL). Treatment with BPL is purportedly effective, with limited influence on SARS-CoV-2 virion morphology and viral proteins [1,14]. Similarly, heat and BPL have been extensively used for treatment of both patient samples and viral preparations to reduce biosafety risk and assist in the development of diagnostic assays [14,15].…”

“…Treatment with BPL is purportedly effective, with limited influence on SARS-CoV-2 virion morphology and viral proteins [1,14]. Similarly, heat and BPL have been extensively used for treatment of both patient samples and viral preparations to reduce biosafety risk and assist in the development of diagnostic assays [14,15]. Jureka et al demonstrate that 56 • C inactivation requires more than 30 min exposure, while 100 • C incubation is effective within 5 min.…”

A Reliable Indirect ELISA Protocol for Detection of Human Antibodies Directed to SARS-CoV-2 NP Protein

“…In this study, we provided a small-scale laboratory validation of robust COVID-19 anti-NP IgG ELISA. This test platform is known to be compatible with large-scale industrial production [ 40 , 41 ]. It can also be adapted to enable pool testing, which minimizes both cost and time for sample processing [ 40 , 41 ].…”

“…This test platform is known to be compatible with large-scale industrial production [ 40 , 41 ]. It can also be adapted to enable pool testing, which minimizes both cost and time for sample processing [ 40 , 41 ]. However, proper optimization and validation under large-scale pooling conditions are required to maintain the assay reliability.…”