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Original Article

J App Pharm Sci. 2022; 12(12): 72-83

In silico design and expression of anti-E1E2 CHIKV scFv in biotinylated form using Escherichia coli Origami B (DE3) for immunochromatographic detection of the Indonesian Chikungunya variant

Korry Novitriani, Ari Hardianto, Bevi Lidya, Ade Rizki Ridwan Firdaus, Bahti Alisjahbana, Muhammad Yusuf, Nur Akmalia Hidayati, Toto Subroto, Shabarni Gaffar.

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Chikungunya is caused by the Chikungunya virus (CHIKV), which is transmitted to humans via the mosquitoes Aedes aegypti or Aedes albopictus. Infected individuals may develop temporary to permanent paralysis; thus, a rapid and accurate detection method is urgently required. The target antigen for the CHIKV antibody recognition is E1E2 glycoprotein which is involved in the attachment of CHIKV to the host cells. This study aimed to design an antiE1E2 CHIKV single-chain variable fragment (scFv) that specifically recognizes the Indonesian CHIKV variant for expression in a biotinylated form using Escherichia coli Origami B (DE3). The recombinant scFv was then applied to detect the E1E2 CHIKV protein by immunochromatography using a streptavidin-biotin system. The scFv protein structure was designed using the anti-E1E2 antigen-binding fragment template and then docked with African E1E2 CHIKV, and the molecular dynamics were simulated using Amber16. The scFv was expressed in fusion form with the biotin acceptor domain using E. coli Origami B (DE3). Biotinylated scFv with a molecular weight of 30 kDa was successfully expressed as characterized by SDS-PAGE, western blotting, and enzyme-linked immunosorbent assay. The immunochromatographic assay showed that anti-E1E2 CHIKV scFv could specifically recognize the E2 CHIKV protein indicated by the positive stains on the nitrocellulose paper.

Key words: Biotinilated anti E1E2 CHIKV scFv, Escherichia coli Origami B (DE3), immunochromatographic detection, Indonesian Chikungunya variant,

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