Cloning of the heavy chain of fragment antigen binding anti-NS1 from hybridoma cell 71E2 induced by dengue virus on pTA2 vector in Escherichia coli TOP10

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

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Dengue infection is one of the major health problems speading worldwide including in Indonesia. The dengue hemorrhagic fever (DHF) incidence in Indonesia has increased rapidly from 1968 until 2013. However, the case fatality ratio decreased during the same period due to early detection of dengue infection by using monoclonal antibody and non-structural 1 (NS1) protein biomarker. This study was conducted to clone the heavy chain of fragment antigen binding (Fab) using local hybridoma clone 71E2 secreting monoclonal antibody anti-NS1, which was induced by the dengue virus. Polymerase chain reaction was used to amplify the heavy chain gene of Fab and agarose gel electrophoresis was used for visualization. The recombinant plasmid that consists of the heavy chain gene and pTA2 vector was transformed to Escherichia coli TOP10. Polymerase chain reaction, digestion, and agarose gel electrophoresis was then used to confirm the gene insertion in the recombinant plasmid. A band of 550 bp appeared in the PCR product of recombinant plasmid, indicating the heavy chain gene of Fab from clone 71E2 has been inserted into the pTA2 vector successfully.

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
CountryIndonesia
CityDepok
Period30/10/1831/10/18

Keywords

  • Cloning
  • dengue
  • Escherichia coli TOP10
  • heavy chain
  • NS1

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    Rahmani, A. F., Pambudi, S., Puteri, S. A., Subiantistha, T., & Lestari, R. (2019). Cloning of the heavy chain of fragment antigen binding anti-NS1 from hybridoma cell 71E2 induced by dengue virus on pTA2 vector in Escherichia coli TOP10. In T. Mart, D. Triyono, & I. T. Anggraningrum (Eds.), Proceedings of the 4th International Symposium on Current Progress in Mathematics and Sciences, ISCPMS 2018 [020104] (AIP Conference Proceedings; Vol. 2168). American Institute of Physics Inc.. https://doi.org/10.1063/1.5132531