Nicotiana tabacum as a potential platform for the production of recombinant anti-toxoplasma single-chain variable fragment (scFv) antibody

Mohamad Zoolkefli, Fatin Iffah Rasyiqah and Go, Pei See and Tan, Boon Chin and Rejab, Nur Ardiyana and Othman, Rofina Yasmin and Khalid, Norzulaani (2020) Nicotiana tabacum as a potential platform for the production of recombinant anti-toxoplasma single-chain variable fragment (scFv) antibody. Malaysian Journal of Science, 39 (2). pp. 1-26. ISSN 1394-3065, DOI https://doi.org/10.22452/mjs.vol39no2.1.

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Official URL: https://doi.org/10.22452/mjs.vol39no2.1

Abstract

Plant systems have now gained much attention for recombinant therapeutic protein production. In this study, we aimed to evaluate the possibility of producing a single-chain variable fragment antibody (scFv), encoded by the TP60 gene, against Toxoplasmosis disease in Nicotiana tabacum cv. SR1. Toxoplasmosis, caused by infection of a parasitic protozoan known as Toxoplasma gondii, is one of the most prevalent parasitic diseases. Leaf explants of N. tabacum were infected with Agrobacterium tumefaciens strain LBA4404 harbouring binary vector pCAMBIA1304 containing TP60 gene. Bands observed at the predicted size of 914 bp confirmed the presence of TP60 transgene, which was stably integrated into both T0 and T1 tobacco plant genome. FV12-6 transgenic line produced the highest mRNA expression (7-fold) correlated to the highest accumulation of anti- Toxoplasma recombinant scFv antibody (0.52% of the total soluble protein), followed by FV16-10, FV17-7, and FV3-11. Normal growth of the transgenic plants was observed. The segregation analysis of FV3, FV12, and FV16 in T1 generation confirmed the transgene integration within a single locus according to 3:1 Mendelian’s law. These findings indicate the potential of using N. tabacum as a host system to produce TP60 recombinant protein without affecting the normal phenotype of the host plant. © 2020 Malaysian Abstracting and Indexing System. All rights reserved.

Item Type: Article
Funders: Ministry of Higher Education, High Impact Research Programme (UM.C/625/1/HIR/MOHE/SCI/18), Post-graduate Research Grant of University of Malaya (PG204-2016A), Research University (RU006-2017 and ST003-2018) grants
Uncontrolled Keywords: Plant molecular farming; Plant transformation; Recombinant protein; Single-chain variable fragment antibody (scFv); Tobacco; Toxoplasmosis
Subjects: Q Science > QH Natural history
S Agriculture > S Agriculture (General)
Divisions: Deputy Vice Chancellor (Research & Innovation) Office > Centre for Research in Biotechnology for Agriculture
Depositing User: Ms. Juhaida Abd Rahim
Date Deposited: 20 Jan 2021 03:39
Last Modified: 20 Jan 2021 03:39
URI: http://eprints.um.edu.my/id/eprint/25693

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