The invention provides a preparation method and applications of pig spleen transferfactor. According to the preparation method, frozen-breaking, freeze thawing breaking, and centrifugal process are combined for transferfactor production, yield is guaranteed, and at the same time production temperature is reduced, and the activity of the transferfactor can be guaranteed preferably. And in addition, the prepared transferfactor contains no protein, is a small molecular peptide, possesses no pyrogen, is a widely used immunomodulator, is capable of increasing vaccine titer, and can be used for improving animal immunity and improving production performance; the activity is not influenced by pancreatic enzymes; and no toxic reaction is caused. When the pig spleen transferfactor is combined with vaccine, animal immunity can be improved, vaccine titer is increased at the same time, body immune response period is shortened, and antibody lasting time is prolonged.