Document Detail


Improved methods for prepurification and detection of staphylococcal enterotoxin B from cell-free culture filtrate.
MedLine Citation:
PMID:  16080722     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Maria Dainiak; Martin Hedström; Igor Yu Galaev; Bo Mattiasson
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biotechnology progress     Volume:  21     ISSN:  8756-7938     ISO Abbreviation:  Biotechnol. Prog.     Publication Date:    2005 Jul-Aug
Date Detail:
Created Date:  2005-08-05     Completed Date:  2006-01-10     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  8506292     Medline TA:  Biotechnol Prog     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1347-51     Citation Subset:  IM    
Affiliation:
Department of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, Box 124, 221 00, Lund, Sweden.
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MeSH Terms
Descriptor/Qualifier:
Calibration
Cell-Free System*
Enterotoxins / analysis*,  isolation & purification*
Enzyme-Linked Immunosorbent Assay / methods*
Hot Temperature
Hydrogen-Ion Concentration
Immunoglobulin Fab Fragments / chemistry
Immunoglobulin G / metabolism
Papain / chemistry
Chemical
Reg. No./Substance:
0/Enterotoxins; 0/Immunoglobulin Fab Fragments; 0/Immunoglobulin G; 39424-53-8/enterotoxin B, staphylococcal; EC 3.4.22.2/Papain

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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