Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Development of a Cost-Effective Process for the Heterologous Production of SARS-CoV-2 Spike Receptor Binding Domain (RBD) Using Pichia pastoris in Stirred-Tank Bioreactor

Diego Noseda
,
Cecilia D’Alessio
,
Javier Santos
ORCID logo ,
Tommy Idrovo-Hidalgo
ORCID logo ,
Maria Florencia Pignataro
ORCID logo ,
Diana E Wetzler
,
Hernán Gentilli
ORCID logo ,
Alejandro D Nadra
ORCID logo ,
Ernesto Roman
,
Carlos Paván
,
Lucas AM Ruberto
*
Version 1 : Received: 24 April 2023 / Approved: 24 April 2023 / Online: 24 April 2023 (04:33:55 CEST)

A peer-reviewed article of this Preprint also exists.

Noseda, D.G.; D’Alessio, C.; Santos, J.; Idrovo-Hidalgo, T.; Pignataro, F.; Wetzler, D.E.; Gentili, H.; Nadra, A.D.; Roman, E.; Paván, C.; Ruberto, L.A.M. Development of a Cost-Effective Process for the Heterologous Production of SARS-CoV-2 Spike Receptor Binding Domain Using Pichia pastoris in Stirred-Tank Bioreactor. Fermentation 2023, 9, 497. Noseda, D.G.; D’Alessio, C.; Santos, J.; Idrovo-Hidalgo, T.; Pignataro, F.; Wetzler, D.E.; Gentili, H.; Nadra, A.D.; Roman, E.; Paván, C.; Ruberto, L.A.M. Development of a Cost-Effective Process for the Heterologous Production of SARS-CoV-2 Spike Receptor Binding Domain Using Pichia pastoris in Stirred-Tank Bioreactor. Fermentation 2023, 9, 497.

Abstract

SARS-CoV-2 was identified as the pathogenic agent causing the COVID-19 pandemic. Among the proteins codified by this virus, the Spike is one of the most external and exposed. A fragment of the Spike protein, named the Receptor Binding Protein (RBD) interacts with the ACE2 receptors of human cells, allowing the entrance of the viruses. RBD has been proposed as an interesting protein for the development of diagnosis tools, treatment and prevention of the disease. In this work, a method for recombinant RBD production using Pichia pastoris as a cell factory in a Stirred Tank Bioreactor (SRTB) up to 7 L was developed. Using a basal saline medium with glycerol, methanol and compressed air in a four stages procedure, around 500 mg/L of raw yRBD (RBD produced by yeasts) and 206 mg/L of purified (>95%) RBD were obtained. Thereby, the proposed method represents a feasible, simple, scalable and cheap procedure for the obtention of RBD for diagnosis kits and vaccines formulation

Keywords

SARS-CoV-2; RBD; ANTIGEN; STBR; PICHIA PASTORIS; BIOREACTOR

Subject

Biology and Life Sciences, Biology and Biotechnology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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