The first report on the sortase-mediated display of bioactive protein A from Staphylococcus aureus (SpA) on the surface of the vegetative form of Bacillus subtilis

Abstract Protein A (SpA) is one of the most important Staphylococcus aureus cell wall proteins. It includes five immunoglobulin (Ig)-binding domains which can bind to immune complexes through the Fc region of immunoglobulins. The binding of SpA to the polymeric supports can be used to prepare affini...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Samira Ghaedmohammadi, Gholamreza Ahmadian
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	Surface display Bacillus subtilis Protein A Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/985cba950d63484398a8c69d02153255
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:985cba950d63484398a8c69d02153255
record_format	dspace
spelling	oai:doaj.org-article:985cba950d63484398a8c69d021532552021-11-21T12:40:40ZThe first report on the sortase-mediated display of bioactive protein A from Staphylococcus aureus (SpA) on the surface of the vegetative form of Bacillus subtilis10.1186/s12934-021-01701-41475-2859https://doaj.org/article/985cba950d63484398a8c69d021532552021-11-01T00:00:00Zhttps://doi.org/10.1186/s12934-021-01701-4https://doaj.org/toc/1475-2859Abstract Protein A (SpA) is one of the most important Staphylococcus aureus cell wall proteins. It includes five immunoglobulin (Ig)-binding domains which can bind to immune complexes through the Fc region of immunoglobulins. The binding of SpA to the polymeric supports can be used to prepare affinity chromatography resins, which are useful for immunoprecipitation (IP) of antibodies. Protein A is also used to purify many anti-cancer antibodies. In this study, SpA was displayed on the surface of Bacillus subtilis cells using a sortase-mediated system to display the target protein to the B. subtilis cell wall. A series of plasmids consisting of cassettes for cell wall-directed protein A as well as negative controls were constructed and transformed into B. subtilis WASD (wprA sigD) cells. SDS-PAGE, western blot, flow cytometry, functional IgG purification assay, and a modified ELISA assay were used to confirm the surface display of SpA and evaluate its function. Semi-quantitative ELISA results showed that the binding capacity of lyophilized Bs-SpA is 100 μg IgG from rabbit serum per 1 mg of cells under optimal experimental conditions. Low production costs, optimal performance, and the use of a harmless strain compared to a similar commercial product predict the possible use of SpA immobilization technology in the future.Samira GhaedmohammadiGholamreza AhmadianBMCarticleSurface displayBacillus subtilisProtein AMicrobiologyQR1-502ENMicrobial Cell Factories, Vol 20, Iss 1, Pp 1-11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Surface display Bacillus subtilis Protein A Microbiology QR1-502
spellingShingle	Surface display Bacillus subtilis Protein A Microbiology QR1-502 Samira Ghaedmohammadi Gholamreza Ahmadian The first report on the sortase-mediated display of bioactive protein A from Staphylococcus aureus (SpA) on the surface of the vegetative form of Bacillus subtilis
description	Abstract Protein A (SpA) is one of the most important Staphylococcus aureus cell wall proteins. It includes five immunoglobulin (Ig)-binding domains which can bind to immune complexes through the Fc region of immunoglobulins. The binding of SpA to the polymeric supports can be used to prepare affinity chromatography resins, which are useful for immunoprecipitation (IP) of antibodies. Protein A is also used to purify many anti-cancer antibodies. In this study, SpA was displayed on the surface of Bacillus subtilis cells using a sortase-mediated system to display the target protein to the B. subtilis cell wall. A series of plasmids consisting of cassettes for cell wall-directed protein A as well as negative controls were constructed and transformed into B. subtilis WASD (wprA sigD) cells. SDS-PAGE, western blot, flow cytometry, functional IgG purification assay, and a modified ELISA assay were used to confirm the surface display of SpA and evaluate its function. Semi-quantitative ELISA results showed that the binding capacity of lyophilized Bs-SpA is 100 μg IgG from rabbit serum per 1 mg of cells under optimal experimental conditions. Low production costs, optimal performance, and the use of a harmless strain compared to a similar commercial product predict the possible use of SpA immobilization technology in the future.
format	article
author	Samira Ghaedmohammadi Gholamreza Ahmadian
author_facet	Samira Ghaedmohammadi Gholamreza Ahmadian
author_sort	Samira Ghaedmohammadi
title	The first report on the sortase-mediated display of bioactive protein A from Staphylococcus aureus (SpA) on the surface of the vegetative form of Bacillus subtilis
title_short	The first report on the sortase-mediated display of bioactive protein A from Staphylococcus aureus (SpA) on the surface of the vegetative form of Bacillus subtilis
title_full	The first report on the sortase-mediated display of bioactive protein A from Staphylococcus aureus (SpA) on the surface of the vegetative form of Bacillus subtilis
title_fullStr	The first report on the sortase-mediated display of bioactive protein A from Staphylococcus aureus (SpA) on the surface of the vegetative form of Bacillus subtilis
title_full_unstemmed	The first report on the sortase-mediated display of bioactive protein A from Staphylococcus aureus (SpA) on the surface of the vegetative form of Bacillus subtilis
title_sort	first report on the sortase-mediated display of bioactive protein a from staphylococcus aureus (spa) on the surface of the vegetative form of bacillus subtilis
publisher	BMC
publishDate	2021
url	https://doaj.org/article/985cba950d63484398a8c69d02153255
work_keys_str_mv	AT samiraghaedmohammadi thefirstreportonthesortasemediateddisplayofbioactiveproteinafromstaphylococcusaureusspaonthesurfaceofthevegetativeformofbacillussubtilis AT gholamrezaahmadian thefirstreportonthesortasemediateddisplayofbioactiveproteinafromstaphylococcusaureusspaonthesurfaceofthevegetativeformofbacillussubtilis AT samiraghaedmohammadi firstreportonthesortasemediateddisplayofbioactiveproteinafromstaphylococcusaureusspaonthesurfaceofthevegetativeformofbacillussubtilis AT gholamrezaahmadian firstreportonthesortasemediateddisplayofbioactiveproteinafromstaphylococcusaureusspaonthesurfaceofthevegetativeformofbacillussubtilis
_version_	1718418900590264320

The first report on the sortase-mediated display of bioactive protein A from Staphylococcus aureus (SpA) on the surface of the vegetative form of Bacillus subtilis

Ejemplares similares