Site-specific albumination of a therapeutic protein with multi-subunit to prolong activity in vivo.

摘要: Albumin fusion/conjugation (albumination) has been an effective method to prolong in vivo half-life of therapeutic proteins. However, its broader application proteins with complex folding pathway or multi-subunit is restricted by incorrect folding, poor expression, heterogeneity, and loss native activity the linked albumin. We hypothesized that site-specific conjugation albumin a permissive site target protein will expand utilities as extender structure. show here genetic incorporation non-natural amino acid (NNAA) followed chemoselective vivo. Urate oxidase (Uox), enzyme for treatment hyperuricemia, homotetramer multiple surface lysines, limiting conventional approaches albumination. Incorporation p-azido-l-phenylalanine into two predetermined positions Uox allowed linkage dibenzocyclooctyne-derivatized human serum (HSA) through strain-promoted azide-alkyne cycloaddition (SPAAC). The bio-orthogonality SPAAC resulted production chemically well-defined conjugate, Uox-HSA, retained enzymatic activity. Uox-HSA had 8.8 h mice, while wild-type 1.3 h. AUC increased 5.5-fold (1657 vs. 303 mU/mL x h). These results clearly demonstrated albumination led prolonged Site-specific enabled NNAA orthogonal chemistry demonstrates promise development long-acting therapeutics high potency safety.