The disclosure provides trispecific and/or trivalent binding proteins comprising four polypeptide chains that form three antigen binding sites that specifically bind one or more target proteins, wherein a first pair of polypeptides forming the binding protein possess dual variable domains having a cross-over orientation and wherein a second pair of polypeptides forming the binding protein possess a single variable domain. The disclosure also provides methods for making trispecific and/or trivalent binding proteins and uses of such binding proteins.本發明提供包含4條多肽鏈的三特异性和/或三價結合蛋白，所述4條多肽鏈形成特异性結合一個或多個靶蛋白的3個抗原結合位點，其中形成該結合蛋白的第一對多肽具有雙重可變域，其具有交叉(cross-over)取向，且其中形成該結合蛋白的第二對多肽具有單一可變域。本文還提供用于製備三特异性和/或三價結合蛋白的方法和此類結合蛋白的用途。
