The present disclosure relates to the field of biological technologies, and discloses a method for expressing and preparing a polyvalent multi-specific antibody and an immune hybrid protein. In the present disclosure, by using the characteristics of protein Intein, a novel method for preparing a polyvalent specific antibody, a hybrid immune protein having an antibody fused to a cytokine, an immunotoxin having an antibody fused to a toxin, or an immune hybrid protein having an antibody fused to other active proteins is designed and developed. In the present disclosure, each portion of a hybrid protein is respectively expressed in a suitable prokaryotic or eukaryotic cell system, separated and purified by high-performance affinity chromatography, and then spliced in vitro by trans-splicing mediated by the intein, to prepare a polyvalent specific antibody and an immune hybrid protein. The method has high production efficiency, and wide scope of application, and facilitates the separation and purification of the products. The present disclosure provides a novel method for preparing biological drugs for directionally attacking cancers or other diseases.
