Vaccination Against Hydatidosis: Molecular Cloning and Optimal Expression of the EG95NC− Recombinant Antigen in Escherichia coli

Cystic echinococcosis (CE) is a widely distributed zoonosis that is highly endemic in the Mediterranean basin. The disease represents a serious public health threat and causes economic losses. The parasite life-cycle involves dogs and ruminants as definitive and intermediate hosts; humans are accide...

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Bibliographic Details
Published in:The Protein Journal 2017-12, Vol.36 (6), p.472-477
Main Authors: Jazouli, M., Lightowlers, M., Gauci, C. G., Tadlaoui, K., Belmlih, A., Ennaji, M. M., Elharrak, M.
Format: Article
Language:English
Subjects:
Affinity chromatography
Animal Anatomy
Antigens
Antigens, Helminth - chemistry
Antigens, Helminth - genetics
Antigens, Helminth - isolation & purification
Antigens, Helminth - metabolism
Biochemistry
Bioorganic Chemistry
Cell culture
Cell density
Chemistry
Chemistry and Materials Science
Clinical trials
Cloning
Cloning, Molecular
Disease transmission
E coli
Echinococcosis
Economic impact
Escherichia coli
Escherichia coli - genetics
Formulations
Gene expression
GST antigen
Helminth Proteins - chemistry
Helminth Proteins - genetics
Helminth Proteins - isolation & purification
Helminth Proteins - metabolism
Histology
Immunization
Liquid chromatography
Molecular biology
Morphology
Organic Chemistry
Protein purification
Proteins
Public health
Recombinant Proteins - chemistry
Recombinant Proteins - genetics
Recombinant Proteins - isolation & purification
Recombinant Proteins - metabolism
Sheep
Vaccination
Vaccines
Vaccines - chemistry
Vaccines - genetics
Vaccines - isolation & purification
Vaccines - metabolism
Zoonoses
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Summary:Cystic echinococcosis (CE) is a widely distributed zoonosis that is highly endemic in the Mediterranean basin. The disease represents a serious public health threat and causes economic losses. The parasite life-cycle involves dogs and ruminants as definitive and intermediate hosts; humans are accidently infected, causing serious clinical issues. Vaccination of ruminants and dog treatments represent the most efficient measures to prevent parasite transmission. The recombinant protein vaccine, EG95, has been used successfully in sheep vaccine trials against CE in several countries. In this study, we expressed the modified antigen, EG95NC-GST, in Escherichia coli for use as a vaccine against Echinococcus granulosus in ruminants. We tested three different media formulations for E. coli culture and established for each culture conditions for optimal levels of soluble EG95 expression. The results demonstrate that SOC and TB media provided high yields in cell density and EG95 protein expression. Purification of the recombinant protein with affinity chromatography (using FPLC) was also performed to increase the purity of the EG95NC − -GST antigen.
ISSN:1572-3887
1573-4943
1875-8355
DOI:10.1007/s10930-017-9742-x