Anti-Her2/VEGFR2/CD3(Peking University Shenzhen Graduate School)

Engineered antibody therapies have significantly propelled the advancement of tumor immunotherapy. However, a considerable proportion of patients fail to respond to treatment or experience relapses following an initial positive response. Moreover, recurrence rates have increased due to antigen escape or downregulation. To address this challenge, novel strategies to engineer trispecific antibodies (tsAbs) have been developed in recent years. Here, we describe the molecular procedures of construction, expression, and purification of optimal tsAbs [specifically, Her2/VEGFR2/CD3 (SO) and CD19/CD22/CD3 (CC) tsAbs]. Also, we detail the experimental methods of functional validation of optimal tsAbs, including immunofluorescence analysis, cytokine release detection, and in vitro cytotoxicity assay and in vivo efficacy studies.