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Plasmodium vivax: comparison of immunogenicity among proteins expressed in the cell-free systems of Escherichia coli and wheat germ by suspension array assays

BACKGROUND: In vitro cell-free systems for protein expression with extracts from prokaryotic (Escherichia coli) or eukaryotic (wheat germ) cells coupled to solid matrices have offered a valid approach for antigen discovery in malaria research. However, no comparative analysis of both systems is pres...

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Autores principales: Rui, Edmilson, Fernandez-Becerra, Carmen, Takeo, Satoru, Sanz, Sergi, Lacerda, Marcus VG, Tsuboi, Takafumi, del Portillo, Hernando A
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3224337/
https://www.ncbi.nlm.nih.gov/pubmed/21756349
http://dx.doi.org/10.1186/1475-2875-10-192
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author Rui, Edmilson
Fernandez-Becerra, Carmen
Takeo, Satoru
Sanz, Sergi
Lacerda, Marcus VG
Tsuboi, Takafumi
del Portillo, Hernando A
author_facet Rui, Edmilson
Fernandez-Becerra, Carmen
Takeo, Satoru
Sanz, Sergi
Lacerda, Marcus VG
Tsuboi, Takafumi
del Portillo, Hernando A
author_sort Rui, Edmilson
collection PubMed
description BACKGROUND: In vitro cell-free systems for protein expression with extracts from prokaryotic (Escherichia coli) or eukaryotic (wheat germ) cells coupled to solid matrices have offered a valid approach for antigen discovery in malaria research. However, no comparative analysis of both systems is presently available nor the usage of suspension array technologies, which offer nearly solution phase kinetics. METHODS: Five Plasmodium vivax antigens representing leading vaccine candidates were expressed in the E. coli and wheat germ cell-free systems at a 50 μl scale. Products were affinity purified in a single-step and coupled to luminex beads to measure antibody reactivity of human immune sera. RESULTS: Both systems readily produced detectable proteins; proteins produced in wheat germ, however, were mostly soluble and intact as opposed to proteins produced in E. coli, which remained mostly insoluble and highly degraded. Noticeably, wheat germ proteins were recognized in significantly higher numbers by sera of P. vivax patients than identical proteins produced in E. coli. CONCLUSIONS: The wheat germ cell-free system offers the possibility of expressing soluble P. vivax proteins in a small-scale for antigen discovery and immuno-epidemiological studies using suspension array technology.
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spelling pubmed-32243372011-11-27 Plasmodium vivax: comparison of immunogenicity among proteins expressed in the cell-free systems of Escherichia coli and wheat germ by suspension array assays Rui, Edmilson Fernandez-Becerra, Carmen Takeo, Satoru Sanz, Sergi Lacerda, Marcus VG Tsuboi, Takafumi del Portillo, Hernando A Malar J Methodology BACKGROUND: In vitro cell-free systems for protein expression with extracts from prokaryotic (Escherichia coli) or eukaryotic (wheat germ) cells coupled to solid matrices have offered a valid approach for antigen discovery in malaria research. However, no comparative analysis of both systems is presently available nor the usage of suspension array technologies, which offer nearly solution phase kinetics. METHODS: Five Plasmodium vivax antigens representing leading vaccine candidates were expressed in the E. coli and wheat germ cell-free systems at a 50 μl scale. Products were affinity purified in a single-step and coupled to luminex beads to measure antibody reactivity of human immune sera. RESULTS: Both systems readily produced detectable proteins; proteins produced in wheat germ, however, were mostly soluble and intact as opposed to proteins produced in E. coli, which remained mostly insoluble and highly degraded. Noticeably, wheat germ proteins were recognized in significantly higher numbers by sera of P. vivax patients than identical proteins produced in E. coli. CONCLUSIONS: The wheat germ cell-free system offers the possibility of expressing soluble P. vivax proteins in a small-scale for antigen discovery and immuno-epidemiological studies using suspension array technology. BioMed Central 2011-07-14 /pmc/articles/PMC3224337/ /pubmed/21756349 http://dx.doi.org/10.1186/1475-2875-10-192 Text en Copyright ©2011 Rui et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methodology
Rui, Edmilson
Fernandez-Becerra, Carmen
Takeo, Satoru
Sanz, Sergi
Lacerda, Marcus VG
Tsuboi, Takafumi
del Portillo, Hernando A
Plasmodium vivax: comparison of immunogenicity among proteins expressed in the cell-free systems of Escherichia coli and wheat germ by suspension array assays
title Plasmodium vivax: comparison of immunogenicity among proteins expressed in the cell-free systems of Escherichia coli and wheat germ by suspension array assays
title_full Plasmodium vivax: comparison of immunogenicity among proteins expressed in the cell-free systems of Escherichia coli and wheat germ by suspension array assays
title_fullStr Plasmodium vivax: comparison of immunogenicity among proteins expressed in the cell-free systems of Escherichia coli and wheat germ by suspension array assays
title_full_unstemmed Plasmodium vivax: comparison of immunogenicity among proteins expressed in the cell-free systems of Escherichia coli and wheat germ by suspension array assays
title_short Plasmodium vivax: comparison of immunogenicity among proteins expressed in the cell-free systems of Escherichia coli and wheat germ by suspension array assays
title_sort plasmodium vivax: comparison of immunogenicity among proteins expressed in the cell-free systems of escherichia coli and wheat germ by suspension array assays
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3224337/
https://www.ncbi.nlm.nih.gov/pubmed/21756349
http://dx.doi.org/10.1186/1475-2875-10-192
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