Expression of polyprotein and 3D polymerase protein in Sf9 cells and immunogenicity against enterovirus A71B5 (Thailand strain)
Issued Date
2023-09-01
Resource Type
eISSN
22313354
Scopus ID
2-s2.0-85172364104
Journal Title
Journal of Applied Pharmaceutical Science
Volume
13
Issue
9
Start Page
27
End Page
36
Rights Holder(s)
SCOPUS
Bibliographic Citation
Journal of Applied Pharmaceutical Science Vol.13 No.9 (2023) , 27-36
Suggested Citation
Issaro N., Kongkaew A., Jittmittraphap A., Leaungwutiwong P., Nimlamool W., Takuathung M.N. Expression of polyprotein and 3D polymerase protein in Sf9 cells and immunogenicity against enterovirus A71B5 (Thailand strain). Journal of Applied Pharmaceutical Science Vol.13 No.9 (2023) , 27-36. 36. doi:10.7324/JAPS.2023.93192 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/90318
Title
Expression of polyprotein and 3D polymerase protein in Sf9 cells and immunogenicity against enterovirus A71B5 (Thailand strain)
Other Contributor(s)
Abstract
Enterovirus 71 B5 subgenotype (EV-A71B5) is a primary human pathogen of hand, foot, and mouth disease. In this study, we aimed to prepare a novel recombinant protein polyprotein (P1) and 3D polymerase (3Dpol) protein of EV-A71B5 and determine mice immunogenicity and neutralization activity against EV-A71 of the B5 subgenotype commonly found in Thailand. Using a dual promoter system and investigating its expression in Sf9 cells, we constructed a novel recombinant protein containing P1 and 3Dpol protein. The purified P1-3Dpol was observed by western blotting and transmission electron microscopy (TEM) to determine the particle size. Furthermore, we determined the immunogenicity and neutralization activity against EV-A71 of the B5 subgenotype using concatenation of Bagg and Albino (BALB/c) mice. The results revealed that P1-3Dpol was expressed in Sf9 cells. We used TEM to visualize the particle size of P1-3Dpol to be approximately 33 nm. P1-3Dpol had the potential to elicit the production of immunoglobulin G and immunoglobulin M antibodies and the T helper type 1-dominant cytokine interferon-γ after immunizing BALB/c mice and inducing neutralization antibodies against EV-A71B5. Our results demonstrated that Sf9 cells successfully produced P1-3Dpol, which can leverage immune efficacy in BALB/c mice and be used to develop vaccines against the EV-A71B5 strain prevalent in Thailand.