Production, Purification and Characterization of Chicken Egg Yolk Monoclonal Antibody Against Colonization factor antigen -1 of Enterotoxigenic Escherichia coli Causing Diarrhea

Enterotoxigenic Escherichia coli (ETEC) causes diarrhea in both humans and animals. The contaminated food and water are the most common vehicles for ETEC infection. The colonization factor antigen (CFA-1) is a fimbriae protein that promotes adherence of the ETEC strain to the epithelium of the small intestine of the host. In this study IgY proteins were produced against the CFA-1 of ETEC in immunized white leg horn chickens. The collection of antibody was found higher in 25% and 50% ammonium precipitated samples. The isolated CFA-1 protein (0.5 mg/ml) was purified by dialysis (0.42 mg/ml), gel-filtration (0.08 mg/ml), and ion-exchange chromatography (0.065 mg/ml). The CFA-1 protein obtained was submitted to pBLAST which matched with CFA-b with accession no.-AAC41415.1. The CFA-1 gene was cloned in pUC18 vector, and transformed into Escherichia coli DH5α strain. In conclusion, we have successfully purified and characterized monoclonal antibodies (IgY proteins) against CFA-1 of enterotoxigenic E. coli that may be useful for diagnosis of widely occurring Escherichia coli infections.