Purification of St enterotoxin produced by enterotoxigenic Escherichia coli (ETEC) (2001)
- Authors:
- Autor USP: TRABULSI, LUIZ RACHID - ICB
- Unidade: ICB
- Assunto: MICROBIOLOGIA
- Language: Inglês
- Abstract: ETEC strains are important agents of diarrhea in general among children in the developing world and the most important cause of traveler's diarrhea. The diarrhea caused by these organisms is mediated by heat-labile (LT) and heat-stable (ST) enterotoxins. In order to develop policlonal and monoclonal antibodies for standardization of immunological assays, ST toxin was purified. E. coli heat-stable enterotoxin from a strain pathogenic for man was mass produced in a chemically defined medium. The toxin was concentrated and purified by sequentially applying batch adsorption chromatography on Amberlite XAD-2 resin, acetone fractionation, and Sephadex G-25 filtration. Reverse-phase high-performance liquid chromatography (HPLC) was used to purify the toxin further. In all steps the ST activity was determined by the suckling mouse assay, samples were diluted and one drop of 1% Evans blue was added per ml and were injected into the stomachs of infant mice. After 4 hours, fluid accumulation in the intestine was measured. Purification of this toxin was an important step to the project. The immune response is not stimulated by the 2 kDa ST toxin, in order to improve its immunogenicity ST was coupled to mouse serum albumin (MSA) or rabbit serum albumin (RSA) or heat-labile toxin (LT) using a bifunctional cross-linking reagent, 1-ethyl-3-(dimethylaminopyl)carbodiimide (EDAC). These compounds were used as immunizing agents for mouse and rabbits
- Imprenta:
- Publisher: Comissão de Cultura e Extensão Universitária do ICB/USP
- Publisher place: São Paulo
- Date published: 2001
- Source:
- Título do periódico: Resumos
- Conference titles: Congresso do Instituto de Ciências Biomédicas
-
ABNT
MENEZES, C A et al. Purification of St enterotoxin produced by enterotoxigenic Escherichia coli (ETEC). 2001, Anais.. São Paulo: Comissão de Cultura e Extensão Universitária do ICB/USP, 2001. . Acesso em: 19 abr. 2024. -
APA
Menezes, C. A., Lima, F. A., Fermnades, M., Franzolin, m R., Magnoli, F., & Trabulsi, L. R. (2001). Purification of St enterotoxin produced by enterotoxigenic Escherichia coli (ETEC). In Resumos. São Paulo: Comissão de Cultura e Extensão Universitária do ICB/USP. -
NLM
Menezes CA, Lima FA, Fermnades M, Franzolin m R, Magnoli F, Trabulsi LR. Purification of St enterotoxin produced by enterotoxigenic Escherichia coli (ETEC). Resumos. 2001 ;[citado 2024 abr. 19 ] -
Vancouver
Menezes CA, Lima FA, Fermnades M, Franzolin m R, Magnoli F, Trabulsi LR. Purification of St enterotoxin produced by enterotoxigenic Escherichia coli (ETEC). Resumos. 2001 ;[citado 2024 abr. 19 ] - Características dos principais grupos de antibacterianos: espectros de ação e indicadores
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