High efficient production of Pr55gag virus-like particles expressing multiple HIV-1 epitopes, including a gp120 protein derived from an Ugandan HIV-1 isolate of subtype A

L. Buonaguro, F. M. Buonaguro, M. L. Tornesello, D. Mantas, E. Beth-Giraldo, R. Wagner, S. Michelson, M. C. Prevost, H. Wolf, G. Giraldo

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

The main goal of this study was to investigate a novel approach for an efficient and reproducible production of Virus-Like Particles (VLPs) expressing multiple HIV-1 epitopes. The HIV-1 Pr55gag-based VLPs have been produced in a Baculovirus expression system, using a transfer vector able to support the independent expression of different open reading frames (ORFs). In this regard, the gp120 derived from 94UG018 HIV-1A isolate, previously studied in our laboratory, has been packaged into the VLPs together with nef and pol ORFs. In particular, the gp120UG sequence shows a 90% homology in the V3 region compared to African HIV-1 strains of the A-clade. This novel approach is extremely effective for the production of VLPs expressing all the epitopes, as confirmed by Western Blot characterization. Furthermore, the resulting HIV-VLPAs show the expected density (1.14-1.18 g/ml) on a 10-60% sucrose gradient and the morphology of an immature virion at standard transmission electron microscopy. Our results demonstrate that this strategy is highly efficient for expressing a balanced amount of multiple epitopes and their packaging in VLP structures, without affecting the Pr55gag autoassembling capacities. Furthermore, the genetic transposition performed in a modified E. coli represents a methodological improvement, allowing a faster and more reproducible identification of recombinant Baculovirus DNA molecules.

Original languageEnglish
Pages (from-to)35-47
Number of pages13
JournalAntiviral Research
Volume49
Issue number1
DOIs
Publication statusPublished - 2001

Fingerprint

Virion
HIV-1
Epitopes
Proteins
Baculoviridae
Open Reading Frames
HIV
Recombinant DNA
Product Packaging
Transmission Electron Microscopy
Sucrose
Western Blotting
Escherichia coli

Keywords

  • Baculovirus
  • Clade-A
  • Vaccine, HIV-1
  • VLPs

ASJC Scopus subject areas

  • Virology
  • Pharmacology

Cite this

High efficient production of Pr55gag virus-like particles expressing multiple HIV-1 epitopes, including a gp120 protein derived from an Ugandan HIV-1 isolate of subtype A. / Buonaguro, L.; Buonaguro, F. M.; Tornesello, M. L.; Mantas, D.; Beth-Giraldo, E.; Wagner, R.; Michelson, S.; Prevost, M. C.; Wolf, H.; Giraldo, G.

In: Antiviral Research, Vol. 49, No. 1, 2001, p. 35-47.

Research output: Contribution to journalArticle

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