Development of a monoclonal antibody-based co-agglutination test to detect enterotoxigenic Escherichia coli isolated from diarrheic neonatal calves
2007
Varshney, Brajesh C. (Intas Biopharmaceuticals Ltd., Gujarat, India), E-mail: [email protected] | Ponnanna, N.M. (National Dairy Developement Board, Gujarat, India) | Sarkar, Pranati A. (National Dairy Developement Board, Gujarat, India) | Rehman, Pragna (National Dairy Developement Board, Gujarat, India) | Shah, Jigar H. (National Dairy Developement Board, Gujarat, India)
Escherichia coli (E. coli) strains were collected from young diarrheic calves in farms and field. Strains that expressed the K99 (F5) antigen were identified by agglutination tests using reference antibodies to K99 antigen and electron microscopy. The K99 antigen from a selected field strain (SAR-14) was heat-extracted and fractionated on a Sepharose CL-4B column. Further purification was carried out by sodium deoxycholate treatment and/or ion-exchange chromatography. Monoclonal antibodies to purified K99 antigen were produced by the hybridoma technique, and a specific clone, NEK99-5.6.12, was selected for propagation in tissue culture.
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