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Title: Advances for the Hepatitis A virus antigen production using a virus strain with codon frequency optimization adjustments in specific locations
Author: Chavarria-Miró, Gemma
Castellarnau Serra, Montserrat de
Fuentes, Cristina
D'Andrea, Lucia
Pérez-Rodríguez, Francisco Javier
Beguiristain, Nerea
Bosch, Albert
Guix Arnau, Susana
Pintó Solé, Rosa
Keywords: Virus de l'hepatitis A
Hepatitis A virus
Issue Date: 18-Feb-2021
Publisher: Frontiers Media
Abstract: The available cell-adapted hepatitis A virus (HAV) strains show a very slow replication phenotype hampering the affordable production of antigen. A fast-growing strain characterized by the occurrence of mutations in the internal ribosome entry site (IRES), combined with changes in the codon composition has been selected in our laboratory. A characterization of the IRES activity of this fast-growing strain (HM175-HP; HP) vs. its parental strain (HM175; L0) was assessed in two cell substrates used in vaccine production (MRC-5 and Vero cells) compared with the FRhK-4 cell line in which its selection was performed. The HP-derived IRES was significantly more active than the L0-derived IRES in all cells tested and both IRES were more active in the FRhK-4 cells. The translation efficiency of the HP-derived IRES was also much higher than the L0-derived IRES, particularly, in genes with a HP codon usage background. These results correlated with a higher virus production in a shorter time for the HP strain compared to the L0 strain in any of the three cell lines tested, and of both strains in the FRhK-4 cells compared to Vero and MRC-5 cells. The addition of wortmannin resulted in the increase of infectious viruses and antigen in the supernatant of FRhK-4 infected cells, independently of the strain. Finally, the replication of both strains in a clone of FRhK-4 cells adapted to grow with synthetic sera was optimal and again the HP strain showed higher yields.
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It is part of: Frontiers in Microbiology, 2021, vol. 12, num. 255
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ISSN: 1664-302X
Appears in Collections:Articles publicats en revistes (Genètica, Microbiologia i Estadística)

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