Heterologous expression of staphylococcal enterotoxin B (seb) gene for antibody production

Staphylococcal food poisoning (SFP) is caused by the members of superantigen family called staphylococcal enterotoxins (SEs). About 20 different types of SEs are produced by Staphylococcus aureus out of which type A (SEA), B (SEB), C (SEC) and D (SED) are commonly implicated in SFP. Among these, SEB...

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Autores principales:	Vrat Kamboj,Dev, Nema,Vijay, Kumar Pandey,Arun, Kumar Goel,Ajay, Singh,Lokendra
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2006
Materias:	affinity purification biotin fusion ELISA gene cloning SEB western blot
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000500010
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spelling	oai:scielo:S0717-345820060005000102007-01-04Heterologous expression of staphylococcal enterotoxin B (seb) gene for antibody productionVrat Kamboj,DevNema,VijayKumar Pandey,ArunKumar Goel,AjaySingh,Lokendra affinity purification biotin fusion ELISA gene cloning SEB western blot Staphylococcal food poisoning (SFP) is caused by the members of superantigen family called staphylococcal enterotoxins (SEs). About 20 different types of SEs are produced by Staphylococcus aureus out of which type A (SEA), B (SEB), C (SEC) and D (SED) are commonly implicated in SFP. Among these, SEB is the most potent toxin and has also gained the status of biological warfare (BW) agent. Therefore, detection of SEB is of utmost importance. Any immunological detection system for SEB requires specific and sensitive antibodies which inturn depends on the purity of the SEB. In the present investigation, seb gene of S. aureus was cloned and expressed in E. coli along with biotin as fusion partner to facilitate the purification process. The yield of purified recombinant SEB was 13.1 mg/L of culture broth. Biotin tag from the biotinylated toxin was removed by protease cleavage, and both biotinylated and non-biotinylated toxin types were used for raising hyperimmune antiserum. Antisera were also specific for SEB amongst different kinds of food poisoning agents tested by indirect plate ELISA and western blot analysis. The quality of the antisera raised in this study was found superior to the commercially available antiserum. The investigation suggests that construction of recombinant staphylococcal enterotoxin B is a good alternative for production of pure enterotoxin to be used in antibody generation.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.9 n.5 20062006-10-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000500010en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	affinity purification biotin fusion ELISA gene cloning SEB western blot
spellingShingle	affinity purification biotin fusion ELISA gene cloning SEB western blot Vrat Kamboj,Dev Nema,Vijay Kumar Pandey,Arun Kumar Goel,Ajay Singh,Lokendra Heterologous expression of staphylococcal enterotoxin B (seb) gene for antibody production
description	Staphylococcal food poisoning (SFP) is caused by the members of superantigen family called staphylococcal enterotoxins (SEs). About 20 different types of SEs are produced by Staphylococcus aureus out of which type A (SEA), B (SEB), C (SEC) and D (SED) are commonly implicated in SFP. Among these, SEB is the most potent toxin and has also gained the status of biological warfare (BW) agent. Therefore, detection of SEB is of utmost importance. Any immunological detection system for SEB requires specific and sensitive antibodies which inturn depends on the purity of the SEB. In the present investigation, seb gene of S. aureus was cloned and expressed in E. coli along with biotin as fusion partner to facilitate the purification process. The yield of purified recombinant SEB was 13.1 mg/L of culture broth. Biotin tag from the biotinylated toxin was removed by protease cleavage, and both biotinylated and non-biotinylated toxin types were used for raising hyperimmune antiserum. Antisera were also specific for SEB amongst different kinds of food poisoning agents tested by indirect plate ELISA and western blot analysis. The quality of the antisera raised in this study was found superior to the commercially available antiserum. The investigation suggests that construction of recombinant staphylococcal enterotoxin B is a good alternative for production of pure enterotoxin to be used in antibody generation.
author	Vrat Kamboj,Dev Nema,Vijay Kumar Pandey,Arun Kumar Goel,Ajay Singh,Lokendra
author_facet	Vrat Kamboj,Dev Nema,Vijay Kumar Pandey,Arun Kumar Goel,Ajay Singh,Lokendra
author_sort	Vrat Kamboj,Dev
title	Heterologous expression of staphylococcal enterotoxin B (seb) gene for antibody production
title_short	Heterologous expression of staphylococcal enterotoxin B (seb) gene for antibody production
title_full	Heterologous expression of staphylococcal enterotoxin B (seb) gene for antibody production
title_fullStr	Heterologous expression of staphylococcal enterotoxin B (seb) gene for antibody production
title_full_unstemmed	Heterologous expression of staphylococcal enterotoxin B (seb) gene for antibody production
title_sort	heterologous expression of staphylococcal enterotoxin b (seb) gene for antibody production
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2006
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000500010
work_keys_str_mv	AT vratkambojdev heterologousexpressionofstaphylococcalenterotoxinbsebgeneforantibodyproduction AT nemavijay heterologousexpressionofstaphylococcalenterotoxinbsebgeneforantibodyproduction AT kumarpandeyarun heterologousexpressionofstaphylococcalenterotoxinbsebgeneforantibodyproduction AT kumargoelajay heterologousexpressionofstaphylococcalenterotoxinbsebgeneforantibodyproduction AT singhlokendra heterologousexpressionofstaphylococcalenterotoxinbsebgeneforantibodyproduction
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Heterologous expression of staphylococcal enterotoxin B (seb) gene for antibody production

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