Unsunnidha, Lalu and Wasito, Raden W and Setyawan, Erif Maha Nugraha and Warsani, Ziana and Kusumawati, Asmarani (2021) Potential of polylactic-co-glycolic acid (PLGA) for delivery Jembrana disease DNA vaccine Model (pEGFP-C1-tat). Journal of Veterinary Science, 22. pp. 1-15. ISSN 1229845X
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Abstract
Background: The development of a vaccine for Jembrana disease is needed to prevent losses in Indonesia's Bali cattle industry. A DNA vaccine model (pEGFP-C1-tat) that requires a functional delivery system will be developed. Polylactic-co-glycolic acid (PLGA) may have potential as a delivery system for the vaccine model.
Objectives: This study aims to evaluate the in vitro potential of PLGA as a delivery system for pEGFP-C1-tat.
Methods: Consensus and codon optimization for the tat gene was completed using a bioinformatic method, and the product was inserted into a pEGFP-C1 vector. Cloning of the pEGFP-C1-tat was successfully performed, and polymerase chain reaction (PCR) and restriction analysis confirmed DNA isolation. PLGA-pEGFP-C1-tat solutions were prepared
for encapsulated formulation testing, physicochemical characterization, stability testing with DNase I, and cytotoxicity testing. The PLGA-pEGFP-C1-tat solutions were transfected in HeLa cells, and gene expression was observed by fluorescent microscopy and real-time PCR.
Results: The successful acquisition of transformant bacteria was confirmed by PCR. The PLGA:DNA:polyvinyl alcohol ratio formulation with optimal encapsulation was 4%:0.5%:2%,
physicochemical characterization of PLGA revealed a polydispersity index value of 0.246, a particle size of 925 nm, and a zeta potential value of −2.31 mV. PLGA succeeded in protecting pEGFP-C1-tat from enzymatic degradation, and the percentage viability from the cytotoxicity
test of PLGA-pEGFP-C1-tat was 98.03%. The PLGA-pEGFP-C1-tat demonstrated luminescence of the EGFP-tat fusion protein and mRNA transcription was detected.
Conclusions: PLGA has good potential as a delivery system for pEGFP-C1-tat.
Item Type: | Article |
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Uncontrolled Keywords: | Jembrana disease; tat gene; DNA vaccine; delivery system; PLGA |
Subjects: | S Agriculture > SF Animal culture |
Divisions: | Faculty of Veterinary Medicine |
Depositing User: | Erlita Cahyaningtyas Cahyaningtyas |
Date Deposited: | 30 Sep 2024 05:04 |
Last Modified: | 30 Sep 2024 05:04 |
URI: | https://ir.lib.ugm.ac.id/id/eprint/7646 |