Process Characterization and Biophysical Analysis for a Yeast-Expressed Phlebotomus papatasi Salivary Protein (PpSP15) as a Leishmania Vaccine Candidate.

Animals Insect Proteins Leishmaniasis Vaccines Mice Phlebotomus Saccharomyces cerevisiae / genetics Salivary Proteins and Peptides / genetics
analytical biochemistry biopharmaceutical characterization biotechnology chromatography protein(s) vaccine(s)

Journal

Journal of pharmaceutical sciences
ISSN: 1520-6017
Titre abrégé: J Pharm Sci
Pays: United States
ID NLM: 2985195R

Informations de publication

Date de publication:
05 2020
Historique:
received: 11 12 2019
revised: 30 01 2020
accepted: 11 02 2020
pubmed: 20 2 2020
medline: 22 6 2021
entrez: 20 2 2020
Statut: ppublish

Résumé

Cutaneous leishmaniasis is a neglected tropical disease caused by the parasite Leishmania and transmitted by sandflies. It has become a major health problem in many tropical and subtropical countries, especially in regions of conflict and political instability. Currently, there are only limited drug treatments and no available licensed vaccine; thus, the need for more therapeutic interventions remains urgent. Previously, a DNA vaccine encoding a 15 kDa sandfly (Phlebotomus papatasi) salivary protein (PpSP15) and recombinant nonpathogenic Leishmania tarentolae secreting PpSP15 have been shown to induce protective immunity against Leishmania major in mice, demonstrating that PpSP15 is a promising vaccine candidate. In this study, we developed a fermentation process in yeast with a yield of ~1g PpSP15/L and a scalable purification process consisting of only 2 chromatographic purification steps with high binding capacity for PpSP15, suggesting that PpSP15 can be produced economically. The biophysical/biochemical analysis of the purified PpSP15 indicated that the protein was of high purity (>97%) and conformationally stable between pH 4.4 and 9.0. More importantly, the recombinant protein had a defined structure similar to that of the related PdSP15 from Phlebotomus duboscqi, implying the suitability of the yeast expression system for producing a correctly folded PpSP15.

Identifiants

DOI: 10.1016/j.xphs.2020.02.004 PMID: 32070701 PMC: PMC7125844
pubmed: 32070701
pii: S0022-3549(20)30077-0
doi: 10.1016/j.xphs.2020.02.004
pmc: PMC7125844
pii:
doi:

Substances chimiques

Insect Proteins 0
Leishmaniasis Vaccines 0
Salivary Proteins and Peptides 0

Types de publication

Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Langues

eng

Sous-ensembles de citation

IM

Pagination

1673-1680

Informations de copyright

Copyright © 2020 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

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Auteurs

Wen-Hsiang Chen (WH)

Departments of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, One Baylor Plaza, BCM113, Houston, Texas 77030; Texas Children's Hospital Center for Vaccine Development, Baylor College of Medicine, 1102 Bates Street, Houston, Texas 77030. Electronic address: Wen-Hsiang.Chen@bcm.edu.

Mun Peak Nyon (MP)

Departments of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, One Baylor Plaza, BCM113, Houston, Texas 77030; Texas Children's Hospital Center for Vaccine Development, Baylor College of Medicine, 1102 Bates Street, Houston, Texas 77030.

Mohan V Poongavanam (MV)

Departments of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, One Baylor Plaza, BCM113, Houston, Texas 77030; Texas Children's Hospital Center for Vaccine Development, Baylor College of Medicine, 1102 Bates Street, Houston, Texas 77030.

Zhuyun Liu (Z)

Departments of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, One Baylor Plaza, BCM113, Houston, Texas 77030; Texas Children's Hospital Center for Vaccine Development, Baylor College of Medicine, 1102 Bates Street, Houston, Texas 77030.

Amadeo B Biter (AB)

Departments of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, One Baylor Plaza, BCM113, Houston, Texas 77030; Texas Children's Hospital Center for Vaccine Development, Baylor College of Medicine, 1102 Bates Street, Houston, Texas 77030.

Rakhi T Kundu (RT)

Departments of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, One Baylor Plaza, BCM113, Houston, Texas 77030; Texas Children's Hospital Center for Vaccine Development, Baylor College of Medicine, 1102 Bates Street, Houston, Texas 77030.

Ulrich Strych (U)

Departments of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, One Baylor Plaza, BCM113, Houston, Texas 77030; Texas Children's Hospital Center for Vaccine Development, Baylor College of Medicine, 1102 Bates Street, Houston, Texas 77030.

Peter J Hotez (PJ)

Texas Children's Hospital Center for Vaccine Development, Baylor College of Medicine, 1102 Bates Street, Houston, Texas 77030; Departments of Pediatrics and Molecular Virology & Microbiology, National School of Tropical Medicine, Baylor College of Medicine, One Baylor Plaza, BCM113, Houston, Texas 77030; Department of Biology, College of Arts and Sciences, Baylor University, Waco, Texas 76706; James A. Baker III Institute for Public Policy, Rice University, Houston, Texas 77005; Hagler Institute for Advanced Study at Texas A&M University, College Station, Houston, Texas 77843.

Maria Elena Bottazzi (ME)

Texas Children's Hospital Center for Vaccine Development, Baylor College of Medicine, 1102 Bates Street, Houston, Texas 77030; Departments of Pediatrics and Molecular Virology & Microbiology, National School of Tropical Medicine, Baylor College of Medicine, One Baylor Plaza, BCM113, Houston, Texas 77030; Department of Biology, College of Arts and Sciences, Baylor University, Waco, Texas 76706. Electronic address: bottazzi@bcm.edu.

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Classifications MeSH

questionsmedicales.fr › Eucaryotes › Animaux › Process Characterization and Biophysical...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines d'arthropode › Protéines d'insecte › Process Characterization and Biophysical...
questionsmedicales.fr › Mélanges complexes › Produits biologiques › Vaccins › Vaccins antiprotozoaires › Vaccins antileishmaniose › Process Characterization and Biophysical...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Rodentia › Muridae › Murinae › Souris › Process Characterization and Biophysical...
questionsmedicales.fr › Eucaryotes › Animaux › Invertébrés › Arthropodes › Insectes › Pterygota › Neoptera › Holometabola › Diptera › Psychodidae › Phlebotomus › Process Characterization and Biophysical...
questionsmedicales.fr › Eucaryotes › Champignons › Levures › Saccharomyces › Saccharomyces cerevisiae › Process Characterization and Biophysical...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines et peptides salivaires › Process Characterization and Biophysical...