The screen of anti-melanoma single chain antibody by phage display technology

Objective: To produce a single chain fragment variable region (ScFv) of anti-melanoma antibody for diagnosis and therapy of the cancer.Methods:Total RNA was extracted from hybridoma cell, and the genes that encode the variable region of light chain and heavy chain were amplified.Then,linked together to construct an ScFv gene.The ScFv gene was cloned into pCANTAB 5E vector, and then competent cell E.coil.TG1 was transformed. After hyperinfection of helper phage M13K07, we obtained a library of ScFv phage antibody. Melanoma cell line LiBr was used to pan recombinant phage antibody for affinity screen. After three rounds of attaching, washing and amplifying, several clones were selected out randomly and underwent an ELISA test.Results:VH,VL and ScFv were identified as 360, 330 and 750bp long, separately. Ten clones of phage antibodies, which showed high affinity, were obtained from thirty randomly selected clones.Conclusion: Anti-melanoma ScFv was prepared successfully by phage display technology. It lays foundation for diagnosis and therapy of melanoma.