Synthesis of disulfide-bridging trehalose polymers for antibody and Fab conjugation using a bis-sulfone ATRP initiator

Herein is reported a bis-sulfone ATRP initiator capable of inserting into the disulfide bonds of antibodies. Further, we present trehalose methacrylate polymers made with this initiator that effectively stabilize antibodies to heat as conjugates.

C-mannosylation supports folding and enhances stability of thrombospondin repeats

Previous studies demonstrated importance of C-mannosylation for efficient protein secretion. To study its impact on protein folding and stability, we analyzed both C-mannosylated and non-C-mannosylated thrombospondin type 1 repeats (TSRs) of netrin receptor UNC-5. In absence of C-mannosylation, UNC-5 TSRs could only be obtained at low temperature and a significant proportion displayed incorrect intermolecular disulfide bridging, which was hardly observed when C-mannosylated. Glycosylated TSRs exhibited higher resistance to thermal and reductive denaturation processes, and the presence of C-mannoses promoted the oxidative folding of a reduced and denatured TSR in vitro. Molecular dynamics simulations supported the experimental studies and showed that C-mannoses can be involved in intramolecular hydrogen bonding and limit the flexibility of the TSR tryptophan-arginine ladder. We propose that in the endoplasmic reticulum folding process, C-mannoses orient the underlying tryptophan residues and facilitate the formation of the tryptophan-arginine ladder, thereby influencing the positioning of cysteines and disulfide bridging.

Highly homogeneous antibody modification through optimisation of the synthesis and conjugation of functionalised dibromopyridazinediones

Herein we report novel protocols for the generation and application of dibromopyridazinediones, an exciting class of disulfide bridging reagents.