The present invention discloses a preparation method for a poultry vaccine-specific pig spleen transfer factor. The preparation method comprises: selecting poultry vaccine antibody-positive pigs, and slaughtering to obtain pig spleen; carrying out cold preservation; thawing the pig spleen; removing fascia; mincing; homogenizing; filling into a bottle, and storing; carrying out repeated freezing and thawing; carrying out centrifugation; carrying out microfiltration; carrying out ultra-filtration; inactivating; adjusting the pH value; adjusting an osmotic pressure; sterilizing; and testing quality. According to the present invention, in the homogenizing step, the pH value of water for injection is adjusted to 4-6 so as to easily increase yields of ribose and polypeptides; the box type member package tangential flow filtration is adopted, such that the residual dead volume of the system is small, and linear enlargement production is easily achieved; the transfer factor crude product is subjected to tangential flow nano-filtration through a 1-3 KD box type member package to prepare the high concentration transfer factor so as to increase a use effect; phenol red is adopted as a pH value indicator, such that customers easily observe the pH value of the transfer factor; and beta-propiolactone is adopted to replace the traditional formaldehyde to inactivate the transfer factor, and the osmotic pressure adjustment process is added so as to easily achieve transfer factor and vaccine combined immunization.