Cloning And Expression Of An Anti-human Liver Cancer Single-chain Fv Gene

Recent developments in the genetic engineering of antibodies have made it possible to use the engineered antibodies in cancer immunotherapy. However, the immunogenecity of currently used murine-derived antibodies greatly limited their clinical application. In recent years,several kinds of engineered antibodies, such as single chain antibodies(scFv),were constructed in order to minimize the immunogenicity and to facilitate the clinical use. Here we report a study on the cloning and expression of an anti-human liver cancer monoclonal antibody(HAb18) single chain Fv gene as well as the activity assay of the expressed product by FCM.The variable region genes of heavy chain (V_H) and light chain (V_L) were amplified from a mouse anti-human liver cancer hybridoma cell line HAb18 by RT-PCR. The genes were cloned,sequenced and compared with those published in EMBL Genbank. The results showed that they are homologous with the...