Objective To construct single chain variable fragment antibody (scFv) from a mouse monoclonal anti - mucin1 (MUC1) antibody secreting cell lines. Methods Isolation of mRNA of hybridoma cells

synthesis of cDNA

and antibody genotyping was performed using mouse antibody specific primers. Related variable genes were fused with a 9 - amino - acid linker to obtain a scFv

which was characterized on denatured and natural antigen by Western blotting and flow cytometry. Results Variable regions of antibodies heavy and light chain coding genes were isolated form a monoclonal anti - MUC1 hybridoma cell line. ScFv was obtained by the combination of the 2 variable genes

which could be produced in

E.coli

. Western blotting and FACS

analysis showed that the obtained scFv could bind to nature and denatured MUC1 of T47D tumor cells specifically. Conclusion A functional anti - MUC1 scFv is obtained successfully

which can bind to tumour associated MUC1 specifically.