Plasmid collection designed for isotype switching
The pFUSE plasmid collection has been designed to change a monoclonal antibody (mAb) from one isotype to another (IgA, IgD, IgE, IgG and IgM). Therefore, they enable the generation of mAbs with the same antigen affinity but with different effector functions, which can either be increased or decreased ADCC (antibody-dependent cellular cytotoxicity), ADCP (antibody-dependent cellular phagocytosis), or CDC (complement dependent cytotoxicity) activity.
This plasmid collection includes:
- pFUSE-CLIg: encodes the constant region of either the kappa or lambda light chain (CL) from a variety of species.
- pFUSE-CHIg: encodes the constant region of a number of Ig isotype heavy chains (CH) from a variety of species.
- pFUSE-Lucia-CHIg: encodes a Lucia luciferase gene to facilitate the detection and quantification of recombinant antibodies.
- Engineered pFUSE-CHIg: encodes mutant forms of the constant regions from the heavy chain of human IgG1 to modify effector function.
Each plasmid in this collection has a unique multiple cloning site (MCS) upstream of the constant region to facilitate the insertion of the variable region of any given mAb.
When pFUSE-CLIg and pFUSE-CHIg are transfected in combination, they can be used to produce entire antibodies from either fragment antigen-binding (Fab) or single-chain variable fragment (scFv) fragments. The whole recombinant mAb can either be chimeric, humanized or fully human depending on the nature of the variable region.