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

Dainiak, Maria LU ; Hedström, Martin LU ; Galaev, Igor LU and Mattiasson, Bo LU (2005) In Biotechnology Progress 21(4). p.1347-1351
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.... (More)
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. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Biotechnology Progress
volume
21
issue
4
pages
1347 - 1351
publisher
The American Chemical Society
external identifiers
  • wos:000231126800045
  • scopus:23244453489
ISSN
1520-6033
DOI
10.1021/bp050099j
language
English
LU publication?
yes
id
d3eed635-be6d-4994-b6c4-f879943669af (old id 155092)
date added to LUP
2007-07-02 09:34:08
date last changed
2017-01-01 07:05:44
@article{d3eed635-be6d-4994-b6c4-f879943669af,
  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       = {Dainiak, Maria and Hedström, Martin and Galaev, Igor and Mattiasson, Bo},
  issn         = {1520-6033},
  language     = {eng},
  number       = {4},
  pages        = {1347--1351},
  publisher    = {The American Chemical Society},
  series       = {Biotechnology Progress},
  title        = {Improved methods for prepurification and detection of Staphylococcal Enterotoxin B from cell-free culture filtrate},
  url          = {http://dx.doi.org/10.1021/bp050099j},
  volume       = {21},
  year         = {2005},
}