METHODS OF CONVERTING FAB SEQUENCES INTO SINGLE CHAIN ANTIBODY SEQUENCES

摘要

In various embodiments, the present invention provides methods of converting a Fab molecule to a scFv molecule. The methods include amplifying the polynucleotide sequence of the variable light and variable heavy regions of the Fab molecule from the framework 1 to framework 4 sequences; adding one or more restriction endonuclease sites to the amino or carboxyl terminal of the amplified variable region sequence by PCR or ligation; adding one or more linker sequences to the amino or carboxyl terminal of the amplified variable region sequence by PCR or ligation. Additionally, in various embodiments the variable light chain sequence is either amino terminal or carboxyl terminal to the variable heavy chain sequence.