| Research Title |
Production of Recombinant scFv Antibody Specific to
Dengue E Protein by Using a Mammalian Expression System |
| Date of Distribution |
21 June 2024 |
| Conference |
| Title of the Conference |
The 4 th International Conference on Science Technology & Innovation Maejo University, Chiang Mai, Thailand |
| Organiser |
The Faculty of Science, Maejo University, Thailand |
| Conference Place |
Chulabhorn Building, Faculty of Science, Maejo University, Chiang Mai, Thailand |
| Province/State |
Chiang Mai |
| Conference Date |
29 March 2024 |
| To |
29 March 2024 |
| Proceeding Paper |
| Volume |
2022 |
| Issue |
4 |
| Page |
194-203 |
| Editors/edition/publisher |
|
| Abstract |
Dengue is a positive single-stranded RNA virus belonging to the Flaviviridae family.
It causes mosquito-borne dengue disease which is a major public health problem in tropical and
subtropical areas. Mouse anti-flavivirus envelope protein antibody (4G2) is one of the common
monoclonal antibodies used in the method for dengue detection. Nowadays, recombinant techniques
have become popular for protein production. This study aims to construct a single chain fragment
variable (scFv) of dengue E protein-specific antibodies by using a mammalian expression system.
Firstly, DNA sequences of 4G2 VH and VL regions were retrieved from GenBank and confirmed the
complementarity-determining regions by the Kabat method. Two sets of primers including universal
primer for VH and VL amplification and specific primer for VH-linker and linker-VL were used. The cDNA
was synthesized from the total RNA of 4G2 hybridoma cells. The VH-linker and linker-VL encoded 4G2
specific to dengue E protein were successfully amplified by PCR with the correct size of VH-linker at
384 bp and linker-VL at 315 bp respectively. Next, the specific restriction enzymes were used to digest
VH-linker and linker-VL before ligation and insertion into the pFLAG-CMV-3 expression vector.
The sequence of the 4G2 scFv-pFLAG-CMV-3 plasmid was confirmed by sequencing technique.
The plasmid was transfected to express 4G2 scFv protein in HEK293T cells and the express protein
was characterized by SDS-PAGE and western blot. In this study, we succeeded in expressing 4G2 scFv
recombinant protein in mammalian cells. The protein will be used for further development of dengue
virus detection method. |
| Author |
|
| Peer Review Status |
มีผู้ประเมินอิสระ |
| Level of Conference |
นานาชาติ |
| Type of Proceeding |
Full paper |
| Type of Presentation |
Poster |
| Part of thesis |
true |
| Presentation awarding |
true |
| Award Title |
Best Poster Presentation |
| Type of award |
รางวัลด้านวิชาการ วิชาชีพ |
| Organiser |
Faculty of Science Maejo University |
| Date of awarding |
29 มีนาคม 2567 |
| Attach file |
|
| Citation |
0
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Publication
Journal Publication
ไทย