The aim of this experiment was to study the effect of the lipoteichoic acid (LTA) extracted from Clostridium butyricum on the adhesion of C. butyricum and Escherichia coli to HT-29 human intestinal cells. The method of extraction of lipoteichoic acid form C. butyricum by TX114 was evaluated. The purification of the LTA by DEAE-cellulose 52 anion exchange chromatography was also investigated. In addition, the LTA was assayed for its inhibition of the adhesion of C. butyricum and E. coli to HT-29 cells and antimicrobial activity. Our results showed that LTA could be extracted by TX114 and purified by DEAE-cellulose 52 anion exchange chromatography and could inhibit the adhesion of C. butyricum and E. coli to HT-29 cells. This result also revealed that the LTA from C. butyricum could inhibit the adhesion of C. butyricum and E. coli to intestinal cells.Key words: Lipoteichoic acid, Clostridium butyricum, HT-29 cells, adhesion, Escherichia col