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dc.contributor.author
Pavan, María Elisa
dc.contributor.author
Pavan, Esteban E.
dc.contributor.author
Cairo, Fabian Martin
dc.contributor.author
Pettinari, María Julia
dc.date.available
2018-06-13T17:17:42Z
dc.date.issued
2016-01
dc.identifier.citation
Pavan, María Elisa; Pavan, Esteban E.; Cairo, Fabian Martin; Pettinari, María Julia; Expression and refolding of the protective antigen of Bacillus anthracis: A model for high-throughput screening of antigenic recombinant protein refolding; Asociación Argentina de Microbiología; Revista Argentina de Microbiología; 48; 1; 1-2016; 5-14
dc.identifier.issn
0325-7541
dc.identifier.uri
http://hdl.handle.net/11336/48529
dc.description.abstract
Bacillus anthracis protective antigen (PA) is a well known and relevant immunogenicprotein that is the basis for both anthrax vaccines and diagnostic methods. Properly foldedantigenic PA is necessary for these applications. In this study a high level of PA was obtained inrecombinant Escherichia coli. The protein was initially accumulated in inclusion bodies, whichfacilitated its efficient purification by simple washing steps; however, it could not be recognizedby specific antibodies. Refolding conditions were subsequently analyzed in a high-throughputmanner that enabled nearly a hundred different conditions to be tested simultaneously. Therecovery of the ability of PA to be recognized by antibodies was screened by dot blot usinga coefficient that provided a measure of properly refolded protein levels with a high degreeof discrimination. The best refolding conditions resulted in a tenfold increase in the intensityof the dot blot compared to the control. The only refolding additive that consistently yieldedgood results was L-arginine. The statistical analysis identified both cooperative and negativeinteractions between the different refolding additives. The high-throughput approach describedin this study that enabled overproduction, purification and refolding of PA in a simple andstraightforward manner, can be potentially useful for the rapid screening of adequate refoldingconditions for other overexpressed antigenic proteins.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Asociación Argentina de Microbiología
dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
Bacillus Anthracis
dc.subject
Protective Antigen
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Protein Refolding
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High-Throughput Refolding Screening
dc.subject.classification
Otras Ciencias Biológicas
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Ciencias Biológicas
dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
dc.title
Expression and refolding of the protective antigen of Bacillus anthracis: A model for high-throughput screening of antigenic recombinant protein refolding
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2018-06-07T20:04:24Z
dc.identifier.eissn
1851-7617
dc.journal.volume
48
dc.journal.number
1
dc.journal.pagination
5-14
dc.journal.pais
Argentina
dc.journal.ciudad
Ciudad Autónoma de Buenos Aires
dc.description.fil
Fil: Pavan, María Elisa. Biochemiq; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina
dc.description.fil
Fil: Pavan, Esteban E.. Politecnico di Milano; Italia
dc.description.fil
Fil: Cairo, Fabian Martin. Biochemiq; Argentina
dc.description.fil
Fil: Pettinari, María Julia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
dc.journal.title
Revista Argentina de Microbiología
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/https://dx.doi.org/10.1016/j.ram.2015.10.004
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0325754115001509
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://www.redalyc.org/articulo.oa?id=213045299002
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