Abstract: In order to develop a specific egg yolk antibody product that can effectively inhibit the two major targets of enterotoxigenic Escherichia coli fimbriae and enterotoxin, this experiment combined enterotoxigenic Escherichia coli K88,K99,987 P,F18,F41 fimbriae and recombinant enterotoxin STa-LTB-STb（SLS） for the preparation of multivalent antigens and the immunization to laying hens. Water dilution-ammonium sulfate precipitation method was combined with ultrafiltration concentration for the separation and purification of egg yolk antibody; SDS-PAGE and ELISA were used to determine its purity and potency; colloidal gold labeling method and hydrocarbon adsorption test were used to investigate its effect on the hydrophobicity of the bacterial surface, and the adsorption to bacteria and antibacterial effect of specific egg yolk antibody was preliminary explored. The results showed that the process used in this experiment could increase the final purity of the egg yolk antibody to 84.8%,and the peak value of the multivalent egg yolk antibody to each antigen component was higher than or equal to 1∶8×10~3,up to 1∶3.2×10~4,which suggested that the multivalent antigen had good immunogenicity and was suitable for mass production of yolk antibody. Under the transmission electron microscope, it was observed that the binding and adsorption effect of the specific egg yolk antibody on the bacteria was obvious. In the LB liquid medium, the proliferation of the five ETEC strains K88,K99,987 P,F18 and F41 were all significantly inhibited. Hydrocarbon adsorption test showed that the binding effect of egg yolk antibody could significantly increase the hydrophobicity of the bacterial surface, making it prone to detach from the water phase. The results suggested that fimbriae-enterotoxin multivalent specific egg yolk antibody could achieve significant antibacterial effect through efficient and specific adsorption and binding with ETEC bacteria.