Engineering a stable CHO cell line for the expression of a MERS-coronavirus vaccine antigen

Vaccine
Mun Peak NyonPeter J Hotez

Abstract

Middle East respiratory syndrome coronavirus (MERS-CoV) has infected at least 2040 patients and caused 712 deaths since its first appearance in 2012, yet neither pathogen-specific therapeutics nor approved vaccines are available. To address this need, we are developing a subunit recombinant protein vaccine comprising residues 377-588 of the MERS-CoV spike protein receptor-binding domain (RBD), which, when formulated with the AddaVax adjuvant, it induces a significant neutralizing antibody response and protection against MERS-CoV challenge in vaccinated animals. To prepare for the manufacture and first-in-human testing of the vaccine, we have developed a process to stably produce the recombinant MERS S377-588 protein in Chinese hamster ovary (CHO) cells. To accomplish this, we transfected an adherent dihydrofolate reductase-deficient CHO cell line (adCHO) with a plasmid encoding S377-588 fused with the human IgG Fc fragment (S377-588-Fc). We then demonstrated the interleukin-2 signal peptide-directed secretion of the recombinant protein into extracellular milieu. Using a gradually increasing methotrexate (MTX) concentration to 5 μM, we increased protein yield by a factor of 40. The adCHO-expressed S377-588-Fc recombinant protein...Continue Reading

Citations

Jul 31, 2018·Expert Review of Vaccines·Yusen ZhouLanying Du
Apr 3, 2020·Vaccines·Jinyong ZhangQuanming Zou
Jun 28, 2020·Journal of Virology·John P Moore, P J Klasse
Jan 20, 2019·Viruses·Craig Schindewolf, Vineet D Menachery
Apr 9, 2020·Frontiers in Microbiology·Ning WangLanying Du
Jun 21, 2020·Vaccines·Cristiano ConteSusanna Esposito
May 12, 2020·Frontiers in Microbiology·Longping V TseRalph S Baric
May 8, 2020·Biomedicines·Kenneth Lundstrom
Apr 25, 2020·Infectious Diseases and Therapy·Eriko Padron-Regalado
Aug 21, 2019·Frontiers in Microbiology·Chean Yeah YongWen Siang Tan
Aug 14, 2020·Human Vaccines & Immunotherapeutics·Mohamad Hesam ShahrajabianQi Cheng
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Feb 18, 2021·Reviews in Medical Virology·Arash ArashkiaUNKNOWN SARS CoV-2 Rapid Response Team of Pasteur Institute of Iran (PII)
Nov 27, 2020·Frontiers in Immunology·Zhihui LiangBingbing Sun
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Aug 25, 2020·Acta Pharmaceutica Sinica. B·Chunting HeXun Sun

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Datasets Mentioned

BETA
AFS88936.1

Methods Mentioned

BETA
PCR
transfection
circular dichroism
electrophoresis
flow cytometry
ELISA
transgenic
Co-IP
glycosylation
flow

Software Mentioned

GeneScript
GraphPad Prism
CDPro
CDSSTR
CONTIN

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