TITLE

EVALUATION OF A MONOCLONAL ANTIBODIES PANEL DIRECTED AGAINST FIMBRIAL ANTIGENS K88, K99, F41 AND 987P OF ENTEROTOXIGENIC ESCHERICHIA COLI

AUTHOR(S)
Junco, J.; Campal, A.; Miranda, A.; Castro, M.; Arteaga, N.; Casas, S.; León, L.; Alvarez, T.; Sanchez, D.; Fernández, G.; Fontirroche, G.
PUB. DATE
October 2002
SOURCE
Revista VacciMonitor (Vacunología y Temas Afines);Oct2002, Vol. 11 Issue 4, p1
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
Enterotoxigenic Escherichia coli (ETEC) represents a large group of strains bacteria that are toxigenic to small intestine of neonates in a number of species including pigs, cattle, sheep, and humans. It well established that the ability of ETEC to colonize the intestine is due to fimbrial antigens wich are able to the bush borders of enterocytes. K88, K99, 987p and F41 were the most common fimbrial antigen of porcine ETEC. We report here the generation and funtional characterization of a dozen of Mouse Monoclonal Antibodies (Mabs) against these ETEC fimbrial antigens. At present, Mabs are essentially supportive in order to develop ETEC diagnosis systems. Different formulations were used for subcutanea and intraperitonial routes of Balb/c mouse immunization for three times monthly. The donor were selected according to the serum title of antibodies determined by ELISA. The hibridomas were obtained from the fusion of spleen lymphocites and mielomas as estándar methods. Antibodies were purified by ion exchange or affinity in relation with their isotype and they were well characterized by Ouchtherlony, SDS-Page, western blot and ELISA techniques for determining their properties. A dozen of Mouse Monoclonal Antibodies (Mabs) against the most common fimbrial antigens of enterotoxigenic Escherichia coli (ETEC) strains: K88, K99, 987p and F41. An important number of IgM isotype antibodies were obtained but, instead of it, an strong IgG antibodies response were elicited. The major of anti-K88 antibodies recognized "a" region. All Mabs were able of recognize their fimbrial antigens only in a native form. All Mabs showed a high affinity for their antigens. The immunization route seem to be an important rol in the antibodies isotype. "a" region of K88 pili showed a very high immunogenicyte, the major of anti-K88 antibodies recognized it. The Mabs recognized their fimbrial antigens only in native form. Mabs looks to be good to used in ELISA and AuBIODOT TM diagnostic systems.
ACCESSION #
27105026

 

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