production of high quality sars cov 2 antigens impact of bioprocess and storage on CORD-Papers-2022-06-02 (Version 1)

Title: Production of high-quality SARS-CoV-2 antigens: Impact of bioprocess and storage on glycosylation biophysical attributes and ELISA serologic tests performance
Abstract: Serological assays are valuable tools to study SARS-CoV-2 spread and importantly to identify individuals that were already infected and would be potentially immune to a virus reinfection. SARS-CoV-2 Spike protein and its receptor binding domain (RBD) are the antigens with higher potential to develop SARS-CoV-2 serological assays. Moreover structural studies of these antigens are key to understand the molecular basis for Spike interaction with angiotensin converting enzyme 2 receptor hopefully enabling the development of COVID-19 therapeutics. Thus it is urgent that significant amounts of this protein became available at the highest quality. In this study we produced Spike and RBD in two human derived cell hosts: HEK293-E6 and Expi293FTM. We evaluated the impact of different and scalable bioprocessing approaches on Spike and RBD production yields and more importantly on these antigens' quality attributes. Using negative and positive sera collected from human donors we show an excellent performance of the produced antigens assessed in serologic enzyme-linked immunosorbent assay (ELISA) tests as denoted by the high specificity and sensitivity of the test. We show robust Spike productions with final yields of approx. 2 mg/L of culture that were maintained independently of the production scale or cell culture strategy. To the best of our knowledge the final yield of 90 mg/L of culture obtained for RBD production was the highest reported to date. An in-depth characterization of SARS-CoV-2 Spike and RBD proteins was performed namely the antigen's oligomeric state glycosylation profiles and thermal stability during storage. The correlation of these quality attributes with ELISA performance show equivalent reactivity to SARS-CoV-2 positive serum for all Spike and RBD produced and for all storage conditions tested. Overall we provide straightforward protocols to produce high-quality SARS-CoV-2 Spike and RBD antigens that can be easily adapted to both academic and industrial settings; and integrate for the first time studies on the impact of bioprocess with an in-depth characterization of these proteins correlating antigen's glycosylation and biophysical attributes to performance of COVID-19 serologic tests.
Published: 2021
Journal: Biotechnol. bioeng
Author Name: Castro Rute
Author link: https://covid19-data.nist.gov/pid/rest/local/author/castro_rute
Author Name: Nobre Lgia S
Author link: https://covid19-data.nist.gov/pid/rest/local/author/nobre_lgia_s
Author Name: Eleutrio Rute P
Author link: https://covid19-data.nist.gov/pid/rest/local/author/eleutrio_rute_p
Author Name: Thomaz Mnica
Author link: https://covid19-data.nist.gov/pid/rest/local/author/thomaz_mnica
Author Name: Pires Antnio
Author link: https://covid19-data.nist.gov/pid/rest/local/author/pires_antnio
Author Name: Monteiro Sandra M
Author link: https://covid19-data.nist.gov/pid/rest/local/author/monteiro_sandra_m
Author Name: Mendes Snia
Author link: https://covid19-data.nist.gov/pid/rest/local/author/mendes_snia
Author Name: Gomes Ricardo A
Author link: https://covid19-data.nist.gov/pid/rest/local/author/gomes_ricardo_a
Author Name: Clemente Joo J
Author link: https://covid19-data.nist.gov/pid/rest/local/author/clemente_joo_j
Author Name: Sousa Marcos F Q
Author link: https://covid19-data.nist.gov/pid/rest/local/author/sousa_marcos_f_q
Author Name: Pinto Filipe
Author link: https://covid19-data.nist.gov/pid/rest/local/author/pinto_filipe
Author Name: Silva Ana C
Author link: https://covid19-data.nist.gov/pid/rest/local/author/silva_ana_c
Author Name: Freitas Micael C
Author link: https://covid19-data.nist.gov/pid/rest/local/author/freitas_micael_c
Author Name: Lemos Ana R
Author link: https://covid19-data.nist.gov/pid/rest/local/author/lemos_ana_r
Author Name: Akpogheneta Onome
Author link: https://covid19-data.nist.gov/pid/rest/local/author/akpogheneta_onome
Author Name: Kosack Lindsay
Author link: https://covid19-data.nist.gov/pid/rest/local/author/kosack_lindsay
Author Name: Bergman Marie Louise
Author link: https://covid19-data.nist.gov/pid/rest/local/author/bergman_marie_louise
Author Name: Duarte Nadia
Author link: https://covid19-data.nist.gov/pid/rest/local/author/duarte_nadia
Author Name: Matoso Paula
Author link: https://covid19-data.nist.gov/pid/rest/local/author/matoso_paula
Author Name: Costa Jlia
Author link: https://covid19-data.nist.gov/pid/rest/local/author/costa_jlia
Author Name: Bandeiras Tiago M
Author link: https://covid19-data.nist.gov/pid/rest/local/author/bandeiras_tiago_m
Author Name: Gomes Alves Patricia
Author link: https://covid19-data.nist.gov/pid/rest/local/author/gomes_alves_patricia
Author Name: Gonalves Carlos P
Author link: https://covid19-data.nist.gov/pid/rest/local/author/gonalves_carlos_p
Author Name: Demengeot Jocelyne
Author link: https://covid19-data.nist.gov/pid/rest/local/author/demengeot_jocelyne
Author Name: Alves Paula M
Author link: https://covid19-data.nist.gov/pid/rest/local/author/alves_paula_m
license: unk
license_url: [unknown license]
source_x: WHO
source_x_url: https://www.who.int/
who_covidence_id: #1098874
has_full_text: FALSE
G_ID: production_of_high_quality_sars_cov_2_antigens_impact_of_bioprocess_and_storage_on