Cloning of recombinant fab from monoclonal antibody anti-dengue NS1 induced by recombinant CHO-K1 cells into pGEM-T vector

T. Subiantistha, S. Pambudi, A. F. Rahmani, S. A. Puteri, R. Lestari

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review

Abstract

Dengue infection, which is an endemic disease in tropical and subtropical countries, is commonly caused by one of the four serotypes of dengue virus. Indonesia as a tropical country has a high case of dengue infection. The dengue virus infection is usually manifested in a fever, but in severe conditions dengue infections are fatal. In order to minimize fatality rate of dengue infection, early diagnostic tests need to be developed. In this study, we aim to produce recombinant Fab from monoclonal antibody anti-Dengue NS1 (44F) induced by recombinant CHO-K1 cells into pGEM-T vector. The Light Chain (LC) of Fab coded by cDNA isolated from hybridoma cell 44F was amplified using PCR. The LC fragment was cloned into pGEM-T vector and transformed into E. coli TOP 10. Several transformants were selected for screening and confirmed by PCR and restriction enzyme digestion. The results showed that the LC gene sized 650 bp was successfully cloned into pGEM-T vector. Further study should be conducted in order to evaluate the level expression of the light chain gene of the Fab fragment as valuable material in the development of dengue diagnostic kit.

Original languageEnglish
Title of host publicationProceedings of the 4th International Symposium on Current Progress in Mathematics and Sciences, ISCPMS 2018
EditorsTerry Mart, Djoko Triyono, Ivandini T. Anggraningrum
PublisherAmerican Institute of Physics Inc.
ISBN (Electronic)9780735419155
DOIs
Publication statusPublished - 4 Nov 2019
Event4th International Symposium on Current Progress in Mathematics and Sciences 2018, ISCPMS 2018 - Depok, Indonesia
Duration: 30 Oct 201831 Oct 2018

Publication series

NameAIP Conference Proceedings
Volume2168
ISSN (Print)0094-243X
ISSN (Electronic)1551-7616

Conference

Conference4th International Symposium on Current Progress in Mathematics and Sciences 2018, ISCPMS 2018
Country/TerritoryIndonesia
CityDepok
Period30/10/1831/10/18

Keywords

  • 44F hybridoma cell
  • dengue
  • E. coli TOP10
  • light chain
  • pGEM-T

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