DEWI ASTRIANY, - and NUR ASNI SETIANI, - and ASAKO YAMAGUCHI, - (2022) Construction of Single-Chain Fragment Variable (ScFv) Anti NS1 Dengue Virus PCR Primers for Phage Display Library Preparation. In: The 5th International Seminar on Pharmaceutical Sciences and Technology (ISPST), Universitas Padjadjaran, 18-20 Oktober 2022, Fakultas Farmasi Universitas Padjadjaran.
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Construction of Single-Chain Fragment Variable (ScFv) Anti NS1 Dengue Virus PCR Primers for Phage Display Library Preparation_ISPST 2022_Dewi Astriany.pdf Download (708kB) |
Abstract
Dengue is an infectious disease that continues to be a public health problem worldwide. The clinical manifestations of dengue infection are difficult to distinguish from other infectious diseases. Rapid and accurate diagnostic tests for dengue virus infection are needed to confirm the disease and treat the patient appropriately. NS1 is the most immunogenic and conserved glycoprotein secreted into the bloodstream1. Therefore, the dengue virus NS1 antigen assay has been identified as one of the specific markers in laboratory diagnostic tests that can be used to detect primary or secondary dengue infection at an early stage. In this study, we designed PCR primers for amplification of VH and VL domains coding for scFv anti NS1. A primary design of scFv anti NS1 dengue virus has been carried out from available genomic databases using bioinformatics analysis which can be used to create a phage display scFv library2,3. The ScFv gene sequence was amplified by PCR with specific primers incorporating NcoI and XhoI restriction sites for directional cloning into a phagemid vector pComb3d-F expression vector4. Cultures of E. coli DH5 were transformed with the pComb3d-F derivatives and were then induced with 0.1 mM isopropyl-d-thiogalactopyranoside (IPTG). The phage display technique was used to generate a panel of the single-chain variable fragment (scFv) antibodies specific for the NS1 protein of the dengue virus. This primer pair has been successfully tested and can be used to produce an scFv phage library, characterize antibodies, and/or design recombinant antibodies for the model.
| Item Type: | Conference or Workshop Item (Speech) |
|---|---|
| Uncontrolled Keywords: | Dengue Virus, NS1, scFv; phage display library; Single-chain Fragment variable |
| Subjects: | Q Science > Q Science (General) |
| Divisions: | Lembaga Penelitian dan Pengabdian Masyarakat > Publikasi Penelitian |
| Depositing User: | - Dewi Astriany - |
| Date Deposited: | 22 Aug 2024 08:34 |
| Last Modified: | 22 Aug 2024 08:34 |
| URI: | http://repository.stfi.ac.id/id/eprint/975 |
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