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Purification of recombinant SARS-CoV-2 spike, its receptor binding domain, and CR3022 mAb for serological assay
Kang Lan Tee; Philip J Jackson; Joseph M Scarrott; Stephen RP Jaffe; Abayomi O Johnson; Yusuf Johari; Thilo H Pohle; Theo Mozzanino; Joseph Price; James Grinham; Adam Brown; Martin J Nicklin; David C James; Mark J Dickman; Tuck Seng Wong.
Afiliación
  • Kang Lan Tee; The University of Sheffield
  • Philip J Jackson; The University of Sheffield
  • Joseph M Scarrott; The University of Sheffield
  • Stephen RP Jaffe; The University of Sheffield
  • Abayomi O Johnson; The University of Sheffield
  • Yusuf Johari; The University of Sheffield
  • Thilo H Pohle; The University of Sheffield
  • Theo Mozzanino; The University of Sheffield
  • Joseph Price; The University of Sheffield
  • James Grinham; The University of Sheffield
  • Adam Brown; The University of Sheffield
  • Martin J Nicklin; The University of Sheffield
  • David C James; The University of Sheffield
  • Mark J Dickman; The University of Sheffield
  • Tuck Seng Wong; The University of Sheffield
Preprint en En | PREPRINT-BIORXIV | ID: ppbiorxiv-231282
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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Texto completo: 1 Colección: 09-preprints Base de datos: PREPRINT-BIORXIV Tipo de estudio: Diagnostic_studies / Prognostic_studies Idioma: En Año: 2020 Tipo del documento: Preprint
Texto completo: 1 Colección: 09-preprints Base de datos: PREPRINT-BIORXIV Tipo de estudio: Diagnostic_studies / Prognostic_studies Idioma: En Año: 2020 Tipo del documento: Preprint