Characterization of receptors for Escherichia coli 987P using competitive binding assays, thin-layer chromatography and gel filtration chromatography

Abstract

Piglet diarrhea is a serious problem in the swine industry. Among various factors causing piglet diarrhea, enterotoxigenic 'Escherichia coli ' (ETEC) is frequently the primary and sole infectious cause. In this study, ETEC bearing 987P fimbriae were selected as the target organism. In order to facilitate studies on the nature of the interaction between 987P fimbriae and its antibody or its receptor, competitive binding assays including a modified form of enzyme-linked immunosorbent assays (ELISA) were developed for the quantitation of the fimbriae, its antibody and receptor using biotinylated 987P fimbriae as a probe. Two immobilization systems were used. One involved the coating of the plates with antibodies; while in the other receptors were the coating material. The glycolipids in the epithelial cells were extracted with chloroform, methanol, and water to detect and identify the receptors for 987P. The extracted glycolipids were separated by high performance thin-layer chromatography (HPTLC) and gel filtration chromatography. The receptor activity was detected using purified 987P fimbriae. (Abstract shortened by UMI.)