Purification of recombinant SARS-CoV-2 spike, its receptor binding domain, and CR3022 mAb for serological assay
Tee, K. L.; Jackson, P. J.; Scarrott, J. M.; Jaffe, S. R.; Johnson, A. O.; Johari, Y.; Pohle, T. H.; Mozzanino, T.; Price, J.; Grinham, J.; Brown, A.; Nicklin, M. J.; James, D. C.; Dickman, M. J.; Wong, T. S.
Show abstract
Serology testing for COVID-19 is highly attractive because of the relatively short diagnosis time and the ability to test for an active immune response against the SARS-CoV-2. In many types of serology tests, the sensitivity and the specificity are directly influenced by the quality of the antigens manufactured. Protein purification of these recombinantly expressed viral antigens [e.g., spike and its receptor binding domain (RBD)] is an important step in the manufacturing process. Simple and high-capacity protein purification schemes for spike, RBD, and CR3022 mAb, recombinantly expressed in CHO and HEK293 cells, are reported in this article. The schemes consist of an affinity chromatography step and a desalting step. Purified proteins were validated in ELISA-based serological tests. Interestingly, extracellular matrix proteins [most notably heparan sulfate proteoglycan (HSPG)] were co-purified from spike-expressing CHO culture with a long cultivation time. HSPG-spike interaction could play a functional role in the pathology and the pathogenesis of SARS-CoV-2 and other coronaviruses.
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