Molecular engineering improves antigen quality and enables integrated manufacturing of a trivalent subunit vaccine candidate for rotavirus.

Autor:	Dalvie NC; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.; The Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA., Brady JR; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.; The Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA., Crowell LE; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.; The Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA., Tracey MK; The Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA., Biedermann AM; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.; The Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA., Kaur K; Department of Pharmaceutical Chemistry, Vaccine Analytics and Formulation Center, University of Kansas, Lawrence, KS, 66047, USA., Hickey JM; Department of Pharmaceutical Chemistry, Vaccine Analytics and Formulation Center, University of Kansas, Lawrence, KS, 66047, USA., Kristensen DL 2nd; The Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA., Bonnyman AD; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.; The Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA., Rodriguez-Aponte SA; The Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA., Whittaker CA; The Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA., Bok M; Instituto de Virología E Innovaciones Tecnológicas, IVIT, CONICET-INTA, Hurlingham,, Buenos Aires, Argentina., Vega C; Instituto de Virología E Innovaciones Tecnológicas, IVIT, CONICET-INTA, Hurlingham,, Buenos Aires, Argentina., Mukhopadhyay TK; Department of Biochemical Engineering, University College London, Gower Street, London, WC1E 6BT, UK., Joshi SB; Department of Pharmaceutical Chemistry, Vaccine Analytics and Formulation Center, University of Kansas, Lawrence, KS, 66047, USA., Volkin DB; Department of Pharmaceutical Chemistry, Vaccine Analytics and Formulation Center, University of Kansas, Lawrence, KS, 66047, USA., Parreño V; Instituto de Virología E Innovaciones Tecnológicas, IVIT, CONICET-INTA, Hurlingham,, Buenos Aires, Argentina., Love KR; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.; The Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA., Love JC; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA. clove@mit.edu.; The Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA. clove@mit.edu.
Jazyk:	angličtina
Zdroj:	Microbial cell factories [Microb Cell Fact] 2021 May 01; Vol. 20 (1), pp. 94. Date of Electronic Publication: 2021 May 01.
DOI:	10.1186/s12934-021-01583-6
Abstrakt:	Background: Vaccines comprising recombinant subunit proteins are well-suited to low-cost and high-volume production for global use. The design of manufacturing processes to produce subunit vaccines depends, however, on the inherent biophysical traits presented by an individual antigen of interest. New candidate antigens typically require developing custom processes for each one and may require unique steps to ensure sufficient yields without product-related variants. Results: We describe a holistic approach for the molecular design of recombinant protein antigens-considering both their manufacturability and antigenicity-informed by bioinformatic analyses such as RNA-seq, ribosome profiling, and sequence-based prediction tools. We demonstrate this approach by engineering the product sequences of a trivalent non-replicating rotavirus vaccine (NRRV) candidate to improve titers and mitigate product variants caused by N-terminal truncation, hypermannosylation, and aggregation. The three engineered NRRV antigens retained their original antigenicity and immunogenicity, while their improved manufacturability enabled concomitant production and purification of all three serotypes in a single, end-to-end perfusion-based process using the biotechnical yeast Komagataella phaffii. Conclusions: This study demonstrates that molecular engineering of subunit antigens using advanced genomic methods can facilitate their manufacturing in continuous production. Such capabilities have potential to lower the cost and volumetric requirements in manufacturing vaccines based on recombinant protein subunits.
Databáze:	MEDLINE
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