Improved methods for prepurification and detection of Staphylococcal Enterotoxin B from cell-free culture filtrate

Maria Dainiak; Martin Hedström; Igor Galaev; Bo Mattiasson

doi:10.1021/bp050099j

Improved methods for prepurification and detection of Staphylococcal Enterotoxin B from cell-free culture filtrate

Maria Dainiak, Martin Hedström, Igor Galaev, Bo Mattiasson

Research output: Contribution to journal › Article › peer-review

Abstract

An improved ELISA method for the detection of Staphylococcal. Enterotoxin B (SEB) in protein A preparations is presented. Fab fragments were obtained by digestion with papain of anti-SEB IgG bound to SEB immobilized on Sepharose 4B. Anti-SEB and peroxidase-labeled Fab fragments from secondary antibodies were successfully used in a modified ELISA of SEB in protein A preparations. SEB-Sepharose was used repeatedly for the production of anti-SEB Fab fragments by papain digestion without loss of affinity. In addition, for the purification of SEB from crude culture filtrates, an initial step utilizing a combined heat and pH treatment for the removal of significant amounts of contaminating proteins without losses of toxin activity is presented. This pretreatment step yielded positive effects in further downstream processing considering both shortened time and an increase in total recovery.

Original language	English
Pages (from-to)	1347-1351
Journal	Biotechnology Progress
Volume	21
Issue number	4
DOIs	https://doi.org/10.1021/bp050099j
Publication status	Published - 2005

Subject classification (UKÄ)

Industrial Biotechnology

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10.1021/bp050099j

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title = "Improved methods for prepurification and detection of Staphylococcal Enterotoxin B from cell-free culture filtrate",

abstract = "An improved ELISA method for the detection of Staphylococcal. Enterotoxin B (SEB) in protein A preparations is presented. Fab fragments were obtained by digestion with papain of anti-SEB IgG bound to SEB immobilized on Sepharose 4B. Anti-SEB and peroxidase-labeled Fab fragments from secondary antibodies were successfully used in a modified ELISA of SEB in protein A preparations. SEB-Sepharose was used repeatedly for the production of anti-SEB Fab fragments by papain digestion without loss of affinity. In addition, for the purification of SEB from crude culture filtrates, an initial step utilizing a combined heat and pH treatment for the removal of significant amounts of contaminating proteins without losses of toxin activity is presented. This pretreatment step yielded positive effects in further downstream processing considering both shortened time and an increase in total recovery.",

author = "Maria Dainiak and Martin Hedstr{\"o}m and Igor Galaev and Bo Mattiasson",

year = "2005",

doi = "10.1021/bp050099j",

language = "English",

volume = "21",

pages = "1347--1351",

journal = "Biotechnology Progress",

issn = "1520-6033",

publisher = "The American Chemical Society (ACS)",

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T1 - Improved methods for prepurification and detection of Staphylococcal Enterotoxin B from cell-free culture filtrate

AU - Dainiak, Maria

AU - Hedström, Martin

AU - Galaev, Igor

AU - Mattiasson, Bo

PY - 2005

Y1 - 2005

N2 - An improved ELISA method for the detection of Staphylococcal. Enterotoxin B (SEB) in protein A preparations is presented. Fab fragments were obtained by digestion with papain of anti-SEB IgG bound to SEB immobilized on Sepharose 4B. Anti-SEB and peroxidase-labeled Fab fragments from secondary antibodies were successfully used in a modified ELISA of SEB in protein A preparations. SEB-Sepharose was used repeatedly for the production of anti-SEB Fab fragments by papain digestion without loss of affinity. In addition, for the purification of SEB from crude culture filtrates, an initial step utilizing a combined heat and pH treatment for the removal of significant amounts of contaminating proteins without losses of toxin activity is presented. This pretreatment step yielded positive effects in further downstream processing considering both shortened time and an increase in total recovery.

AB - An improved ELISA method for the detection of Staphylococcal. Enterotoxin B (SEB) in protein A preparations is presented. Fab fragments were obtained by digestion with papain of anti-SEB IgG bound to SEB immobilized on Sepharose 4B. Anti-SEB and peroxidase-labeled Fab fragments from secondary antibodies were successfully used in a modified ELISA of SEB in protein A preparations. SEB-Sepharose was used repeatedly for the production of anti-SEB Fab fragments by papain digestion without loss of affinity. In addition, for the purification of SEB from crude culture filtrates, an initial step utilizing a combined heat and pH treatment for the removal of significant amounts of contaminating proteins without losses of toxin activity is presented. This pretreatment step yielded positive effects in further downstream processing considering both shortened time and an increase in total recovery.

U2 - 10.1021/bp050099j

DO - 10.1021/bp050099j

M3 - Article

SN - 1520-6033

VL - 21

SP - 1347

EP - 1351

JO - Biotechnology Progress

JF - Biotechnology Progress

IS - 4

ER -

Improved methods for prepurification and detection of Staphylococcal Enterotoxin B from cell-free culture filtrate

Abstract

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