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?:abstract
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The spike protein of the pandemic human Corona virus is essential for virus entry into human cells In fact, most neutralizing antibodies against Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2) are directed against the Virus-surface exposed spike protein, making it the antigen of choice for use in vaccines and diagnostic tests In the current pandemic context, global demand for spike proteins has rapidly increased and could exceed hundreds of grams to kilograms annually Coronavirus spikes are large heavily glycosylated homo-trimeric complexes, with inherent instability The poor manufacturability now threatens the availability of these proteins for vaccines and diagnostic tests Here, we outline scalable, Good Manufacturing Practice (GMP) compliant, and chemically defined processes for the production of two cell-secreted stabilized forms of the trimeric spike proteins (Wuhan and D614G variant) The processes are chemically defined and based on clonal suspension-CHO cell populations and on protein purification via a two-step scalable downstream process The trimeric conformation was confirmed using electron microscopy and HPLC analysis Binding to susceptible cells was shown using a virus-inhibition assay The diagnostic sensitivity and specificity for detection of serum SARS-CoV-2-specific-immunoglobulin molecules was found to exceed that of spike fragments (Spike fragment-1, S1 and Receptor Binding Domain, RBD) The process described here will enable production of sufficient high-quality trimeric spike protein to meet the global demand for SARS-CoV-2 diagnostic tests and potentially vaccines
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