Systematic Production of IgG-Like Bispecific Antibodies via Intein-Mediated Protein Trans-Splicing.

Bispecific antibodies show therapeutic potential; thus, their screening campaign has attracted attention for the development of biotherapeutics. However, many bispecific formats require engineering or fine-tuned purification processes optimized for each molecule, hindering simple screening, especially when impurities are not allowed. In this chapter, we describe a method for producing IgG-like bispecific antibodies via intein-mediated protein trans-splicing, fusing two polypeptide units post-translationally. This method allows the separate expression of two antigen-binding fragments and eliminates the light chain problem. Homobivalent species are not contaminated. In addition, the common purification process finalized by size-exclusion chromatography enables the systematic production of bispecific antibodies suitable for in vitro screening.