Complete Digestion of Mutates Antibodies and Fc-fusion Proteins using FabDELLO®

Performance Activity

Above-hinge digesiton of human IgG antibodies and Fc-fusion proteins, enabling robust characterization of engineered and hinge-mutated biotherapeutics.

Most therapeutic antibodies carry a human IgG1 backbone and the development of bi- and multispecific formats are rapidly increasing. The analysis of such antibodies requires specific enzymes to characterize properties such as monovalent binding, disulfide scrambling and paired Fc glycosylation. Traditional protocols include enzymatic digestion using papain and Lys-C, which require extensive optimizations to minimize overdigestion and obtain sufficient yields. 

FabDELLO digests human IgG1 at a single site above the hinge and generates intact Fab and Fc fragments. To demonstrate the specific activity of FabDELLO, a selection of IgG1 and Fc-fusion proteins were digested and analyzed by non-reduced SDS-PAGE. A complete digestion was achieved for all substrates, including IgG1 with the otherwise difficult LALA and LFLE mutations. The precise and effective performance of the FabDELLO enzyme on this wide range of substrates validates its use for the development of biopharmaceuticals.

Reliable above hinge digestion on a variety of native and mutated molecules

FabDELLO activity on a variety of human IgG1 and Fc-fusion protein substrates. Non-reducing SDS-PAGE analysis of commercial antibodies and Fc-fusion proteins digested with FabDELLO.

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