Optimization of Synthetic Gene Encoding Anti-NS1 Dengue Virus ScFv Expression in Escherichia coli BL21(DE3)

DEWI ASTRIANY, - and UMI BAROROH, - and SYARIF HAMDANI, - (2024) Optimization of Synthetic Gene Encoding Anti-NS1 Dengue Virus ScFv Expression in Escherichia coli BL21(DE3). In: Asian Association of Schools of Pharmacy (AASP) Pharmacy Education Forum 2024, March 6-8, 2024, Bangkok - Thailand.

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Abstract

Dengue is an infectious disease that is still a concern for global health. For the diagnosis of dengue virus infection and the proper course of therapy for the patient, a quick and reliable diagnostic test is essential. The most conserved and immunogenic glycoprotein NS1 antigen is a particular marker in laboratory diagnostic procedures that can be used to identify primary or secondary dengue infection. This research was conducted to express, optimise, and characterise a synthetic gene encoding a single-chain antibody fragment (scFv) against the NS1 protein of the dengue virus fused Biotin Acceptor Domain (BAD) in Escherichia coli BL21 (DE3). The recombinant scFv-BAD fusion protein was expressed in different IPTG concentrations (0.10, 0.55, and 1.00 mM), incubated at certain temperatures (19, 28, and 37°C), and induction time (4, 8, and 12 hours) to promote protein expression in the Luria Bertani medium along with biotin. Optimisation designs were obtained using Central Composite Design and analysed by the Response Surface Method. Using the SDS-PAGE technique, purified recombinant biotinylated scFv was described. The recombinant scFv-BAD fusion protein was partially soluble and partially in the form of inclusion bodies. According to Central Composite Design analysis, 0.67 mM of Isopropyl β-D-Thiogalactopyranoside (IPTG), 4 hours of induction time, and 29.54°C incubation were the optimum conditions for generating 0.4319 mg/mL of recombinant scFv protein on Luria Bertani medium. In conclusion, the level of scFv gene expression in E. coli BL21 (DE3) was significantly influenced by the interaction between the chosen optimisation parameters.

Item Type:	Conference or Workshop Item (Speech)
Uncontrolled Keywords:	Synthetic gene, NS1 antigen, scFv, Escherichia coli, Dengue
Subjects:	Q Science > Q Science (General)
Divisions:	Lembaga Penelitian dan Pengabdian Masyarakat > Publikasi Penelitian
Depositing User:	- Dewi Astriany -
Date Deposited:	18 Aug 2024 14:18
Last Modified:	18 Aug 2024 14:18
URI:	http://repository.stfi.ac.id/id/eprint/853

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