This invention is preparation procedure of the yolk antibody of high biological activity, belonging to the field of bioproduct and industrialized isolation technique. In this invention, we adopt the natural macromolecule gel high -extractive technique and hyperfiltration to extract the yolk-antibody of high biological activity from yolk at an industrialized level, annual output is upto 10 ton. the yolk-antibody content is about 30%-50%, meanwhile we could obtain some by-products like albumen powder, yolk powder, egg oil, vitellin and so on. In the invention high technology like bioimmunology and membrane separation is adopted. The invention is high-tech, high-usage, highly-automated and typical of green food and environmental-friendly.