RP215 single chain fragment variable and single domain recombinant antibodies induce cell cycle arrest at G0/G1 phase in breast cancer.

摘要: Abstract Background Targeted therapy is an attractive approach to avoid the side effects of cancer treatment. Based on antibody-targeted superantigens, single chain variable fragment (scFv) and domain (sdAb) antibodies, characterized by a low molecular weight, immunogenicity high tumor penetration compared monoclonal antibodies (mAb), have been increasingly used in gene-targeted for cancer. In present study, we aimed develop novel recombinant scFv-RP215 sdAb-RP215 based regions RP215 antibody (RP215-mAb) against CA215, pan marker expressed various human tissues, examine their biological activity breast cell lines. Methods The VH VL genes were amplified from hybridoma cells secreting RP215-mAb RT-PCR joined with linker using splicing overlap extension PCR (SOE-PCR) obtain RP215-scFv gene, whereas gene was generate RP215-sdAb. Gene fragments subcloned into pET32a(+) vector Escherichia coli BL21. Western blot, indirect immunofluorescence (IF), ELISA competitive detect immunoreactivity scFv-RP215, sdAb-RP215, RP215-mAb. CCK-8 assay cycle analysis assess function. Results successfully developed CA215. We verified that recognize CA215 surface (MB231, MCF7, MB468, SK-BR-3 BT549) specificity. Our findings also indicate induce arrest at G0/G1 phase cells. Conclusion results showed excellent localize accurately membrane-bound form, suggesting potential as targeting therapy.