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SNAP-Tag Technology: A Useful Tool to Determine Affinity Constants and Other Functional Parameters of Novel Antibody Fragments

: Niesen, J.; Sack, M.; Seidel, M.; Fendel, R.; Barth, S.; Fischer, R.; Stein, C.


Bioconjugate Chemistry 27 (2016), Nr.8, S.1931-1941
ISSN: 1043-1802
Fraunhofer IME ()

Antibody derivatives, such as the single chain fragment variable (scFv), can be developed as diagnostic and therapeutic tools in cancer research, especially in the form of fusion proteins. Such derivatives are easier to produce and modify than monoclonal antibodies (mAbs) and achieve better tissue/tumor penetration. The genetic modification of scFvs is also much more straightforward than the challenging chemical modification of mAbs. Therefore, we constructed two scFvs derived from the approved monoclonal antibodies cetuximab (scFv2112) and panitumumab (scFv1711), both of which are specific for the epidermal growth factor receptor (EGFR), a well-characterized solid tumor antigen. Both scFvs were genetically fused to the SNAP-tag, an engineered version of the human DNA repair enzyme O6-alkylguanine DNA alkyltransferase that allows the covalent coupling of benzylguanine (BG)-modified substrates such as fluorescent dyes.