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Control of Established Colon Cancer Xenografts Using a Novel Humanized Single Chain Antibody-Streptococcal Superantigen Fusion Protein Targeting the 5T4 Oncofetal Antigen

Figure 3

Generation of the scFv5T4::SpeCD203A fusion protein and control reagents.

A) Schematic illustration representing the components of the generated fusion protein constructs. The protein consists of the generated 5T4-targeted humanized single chain variable fragments, VH and VL (grey bar), genetically fused to streptococcal superantigen SpeC (blue bar) either containing an alanine substitution at residue D203 or an additional alanine substitution at residue Y15. All constructs were generated to contain a C-terminal biotin tag. The purified recombinant proteins are shown by SDS-PAGE (panel B), and detected by Western blot analysis by streptavidin-IRDye800 (panel C).

Figure 3