Encapsulation of proteins from Leishmania panamensis into PLGA particles by a single emulsion-solvent evaporation method

J Microbiol Methods. 2019 Jul:162:1-7. doi: 10.1016/j.mimet.2019.05.004. Epub 2019 May 10.

Abstract

The current therapy for the treatment of leishmaniasis is unsatisfactory because it has multiple side effects, and resistance has been reported among the parasites that cause these diseases. Numerous efforts have been made to develop new candidates for vaccines. In recent years, particles of biodegradable polymers have been proposed as vehicles to transport and protect antigens, proteins, drugs and vaccines. In this work, the oil/water (o/w) single emulsion-solvent evaporation technique was used to prepare PLGA biodegradable particles. The encapsulation of two hypothetical proteins from Leishmania panamensis was performed to validate the proposed method. For this validation, different concentrations (50, 100, 150, 200, 250, 500, and 750 μg/ml) of both proteins were encapsulated into PLGA particles, and the particle sizes and shapes were evaluated by optical microscopy and scanning electron microscopy (SEM), respectively. The release of proteins was confirmed by SDS-PAGE and Western blot analyses. The integrity of both proteins was conserved, and they were released from day one until day 15, with a maximum amount of 46 ± 4.25% for the LpanUA.27.1260 protein and 26.19 ± 3.41% for LpanUA.22.1860. Additionally, the protective efficacy of one of these encapsulated proteins was evaluated in vivo using BALB/c mice infected with L. panamensis. Therefore, the encapsulation of proteins is presented here as an excellent alternative to evaluate the antigenicity of proteins from parasites of medical importance such as L. panamensis.

Keywords: Biodegradable polymers; Encapsulated microparticles; Hypothetical proteins; Leishmania; Vaccine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Emulsions
  • Female
  • Leishmania / immunology*
  • Leishmaniasis / therapy*
  • Leishmaniasis Vaccines / administration & dosage
  • Leishmaniasis Vaccines / chemistry*
  • Mice
  • Mice, Inbred BALB C
  • Microspheres
  • Particle Size
  • Polylactic Acid-Polyglycolic Acid Copolymer / chemistry*
  • Protozoan Proteins / administration & dosage
  • Protozoan Proteins / chemistry*
  • Solvents

Substances

  • Emulsions
  • Leishmaniasis Vaccines
  • Protozoan Proteins
  • Solvents
  • Polylactic Acid-Polyglycolic Acid Copolymer