Improved methods for prepurification and detection of Staphylococcal Enterotoxin B from cell-free culture filtrate
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/155092
- author
- Dainiak, Maria LU ; Hedström, Martin LU ; Galaev, Igor LU and Mattiasson, Bo LU
- organization
- publishing date
- 2005
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Biotechnology Progress
- volume
- 21
- issue
- 4
- pages
- 1347 - 1351
- publisher
- The American Chemical Society (ACS)
- 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
- 2016-04-01 16:27:14
- date last changed
- 2022-01-28 19:50:26
@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 (ACS)}},
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}},
doi = {{10.1021/bp050099j}},
volume = {{21}},
year = {{2005}},
}