Expression Of Anti-osteoblastic Sarcoma Single-chain Bifunctional Antibody Fusion Protein And Its Forecast With Networks

Objective: To analyze secondary structure of the fusion protein with bioinformatics of softwares, and to establish a recombinant secretory anti-osteoblastic sarcoma cell single-chain bi-functional antibody gene. Methods: Bioinformatics of softwares was used to analyse structural change of the ScFv-TNF protein. In our works, the sites of enzyme were selected for the sequence of linker, and then DNAssist and ANTHEPROT V5 were used to analyze their sequence, their secondary structure, and their physical and chymic character. A single-chain antibody (ScFv) gene comprising the linked heavy and light chain variable region derived form a monclonal antibody with high specifity and affinity binding to osteoblastic sarcoma cell, TNF was fused in frame with ScFv gene of downstream by PCR. Constructed secretory anti-osteoblastic sarcoma cell single chain bi-function antibody gene. The ScFv-TNF gene was then cloned into an expression vector PLxSN. The vector was identified with enzyme digestions for the correct in section and further confirmed by DNA sequence analysis. By the technology of directional cloning, the sequenced genes were subcloned into corresponding restriction sites of pGEFP-N₃ in turn to generate different...