Objective：To optimize，screen and identify the bispecific c-Met/PD-L1 scFv fusion protein and to detect whether the different combinations of heavy chains and light chains can influence the biological activity of bispecific scFv fragments. Methods：Bioinformatics analysis，technique of gene engineered antibodies were introduced to design，optimize and construct bi-specific scFv fusion proteins. BLItz was used to analyze the affinity of those bi-specific scFv fusion proteins to c-Met and PD-L1，ELISA assay was used to detect their specific binding ability. Results：Bispecific scFv fusion proteins were produced successfully，BLItz and ELISA detection confirmed that the bispecific scFv fusion protein CP1 has higher affinity and specific binding ability. Conclusion：Different combinations of heavy and light chains can affect the affinity and specific binding activity of c-Met/PD-L1 scFv-Fc fusion proteins，CP1 has the strongest affinity and specificity for binding to c-Met and PD-L1 protein，it can be applied to the construction of c-Met/PD-L1 expression vector and subsequent research of c-Met/PD-L1 CAR-T cells.