Process development and scale-up optimization of the SARS-CoV-2 receptor binding domain-based vaccine candidate, RBD219-N1C1.
Jungsoon Lee,Zhuyun Liu,Wen-Hsiang Chen,Junfei Wei,Rakhi Tyagi Kundu,Rakesh Adhikari,Joanne Altieri Rivera,Portia M. Gillespie,Ulrich Strych,Bin Zhan,Peter J. Hotez,Maria Elena Bottazzi +11 more
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In this article, a SARS-CoV-2 RBD219-N1C1 recombinant protein antigen formulated on Alhydrogel® has been shown to elicit a robust neutralizing antibody response against SARS CoV2 pseudovirus in mice.Abstract:
A SARS-CoV-2 RBD219-N1C1 (RBD219-N1C1) recombinant protein antigen formulated on Alhydrogel® has recently been shown to elicit a robust neutralizing antibody response against SARS-CoV-2 pseudovirus in mice. The antigen has been produced under current good manufacturing practices (cGMPs) and is now in clinical testing. Here, we report on process development and scale-up optimization for upstream fermentation and downstream purification of the antigen. This includes production at the 1-L and 5-L scales in the yeast, Pichia pastoris, and the comparison of three different chromatographic purification methods. This culminated in the selection of a process to produce RBD219-N1C1 with a yield of >400 mg per liter of fermentation with >92% purity and >39% target product recovery after purification. In addition, we show the results from analytical studies, including SEC-HPLC, DLS, and an ACE2 receptor binding assay that were performed to characterize the purified proteins to select the best purification process. Finally, we propose an optimized upstream fermentation and downstream purification process that generates quality RBD219-N1C1 protein antigen and is fully scalable at a low cost. KEY POINTS: • Yeast fermentation conditions for a recombinant COVID-19 vaccine were determined. • Three purification protocols for a COVID-19 vaccine antigen were compared. • Reproducibility of a scalable, low-cost process for a COVID-19 vaccine was shown. Graphical abstract.read more
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Scientific rationale for developing potent RBD-based vaccines targeting COVID-19.
Harry Kleanthous,Judith M. Silverman,Karen W. Makar,In-Kyu Yoon,Nicholas Jackson,D. W. Vaughn +5 more
TL;DR: In this paper, the authors reviewed the data supporting the non-inferiority of RBD as a vaccine immunogen compared to full-length S-protein vaccines with respect to humoral and cellular immune responses against both the prototype pandemic SARS-CoV-2 isolate and emerging variants of concern.
Journal ArticleDOI
SARSCoV-2 RBD219-N1C1: A yeast-expressed SARS-CoV-2 recombinant receptor-binding domain candidate vaccine stimulates virus neutralizing antibodies and T-cell immunity in mice.
Jeroen Pollet,Jeroen Pollet,Wen-Hsiang Chen,Wen-Hsiang Chen,Leroy Versteeg,Brian Keegan,Bin Zhan,Bin Zhan,Junfei Wei,Zhuyun Liu,Jungsoon Lee,Rahki Kundu,Rakesh Adhikari,Cristina Poveda,Maria Jose Villar,Ana Carolina de Araujo Leao,Joanne Altieri Rivera,Zoha Momin,Portia M. Gillespie,Jason T. Kimata,Ulrich Strych,Ulrich Strych,Peter J. Hotez,Maria Elena Bottazzi,Maria Elena Bottazzi,Maria Elena Bottazzi +25 more
TL;DR: In this article, a SARS-CoV-2 receptor-binding domain (RBD) protein, expressed at high levels in yeast (Pichia pastoris), was used as a suitable vaccine candidate against COVID-19.
Journal ArticleDOI
Whole Inactivated Virus and Protein-Based COVID-19 Vaccines
TL;DR: The rapid development and deployment of mRNA and adenovirus-vectored vaccines against coronavirus disease 2019 continue to astound the global scientific community, but these vaccine platforms and production approaches have still not achieved global COVID-19 vaccine equity as discussed by the authors .
Journal ArticleDOI
Evaluation of safety and immunogenicity of receptor-binding domain-based COVID-19 vaccine (Corbevax) to select the optimum formulation in open-label, multicentre, and randomised phase-1/2 and phase-2 clinical trials
S. C. Thuluva,Vikram M. Paradkar,S. Gunneri,V. Yerroju,R. Mogulla,K. Turaga,Mahesh Kyasani,Senthil Kumar Manoharan,Guruprasad R. Medigeshi,J. Singh,Heena Shaman,Chandramani Singh,Venkateshwar Rao A +12 more
TL;DR: A randomised Phase-1/2 trial followed by a Phase-2 trial were conducted to assess the safety and immunogenicity of the COVID-19 vaccine Corbevax and select to an optimum formulation as discussed by the authors .
Journal ArticleDOI
Yeast-produced RBD-based recombinant protein vaccines elicit broadly neutralizing antibodies and durable protective immunity against SARS-CoV-2 infection
Jinkai Zang,Yuanfei Zhu,Yu Zhou,Chenjian Gu,Yufang Yi,Shuxia Wang,Shiqi Xu,Gaowei Hu,Shujuan Du,Yannan Yin,Yalei Wang,Yong Yang,Xueyang Zhang,Haikun Wang,Feifei Yin,Feifei Yin,Chao Zhang,Qiang Deng,Youhua Xie,Zhong Huang +19 more
TL;DR: Yeast-derived RBD-monomer possessed functional conformation and was able to elicit a high level of neutralizing antibodies in mice as mentioned in this paper, implying broad-spectrum protection potential of the RBDbased vaccines.
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The EMBL-EBI search and sequence analysis tools APIs in 2019
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TL;DR: The latest improvements made to the frameworks which enhance the interconnectivity between public EMBL-EBI resources and ultimately enhance biological data discoverability, accessibility, interoperability and reusability are described.