GlySERIAS는 Gly4Ser 및 GlyxSery(GS) 및 Polyglycine(G) 링커와 같은 Glycine이 많이 있는 fusion protein linker를 Digestion 할 수 있는  Enzyme입니다. Glycine linker 부분을 특이적으로 digenstion 하여 분석할 수 있으며, 항체의 complex를 잘 이해할 수 있습니다.

   Digestion Workflow

   Digestion of a GS-linked Fusion Protein

Dulaglutide 항체 분석

The flexible GS linker of dulaglutide was digested with GlySERIAS for 1 hour at 37°C under nativeconditions, and the Fc glycans were concurrently hydrolyzed using GlycINATOR to reduce sample complexity. To stop the GlySERIAS reaction, 1 mM ZnCl2 was added. The interchain disulfi de bonds were reduced with 20 mM DTT for 30 minutes at 37°C. The digest was performed in triplicate. a) Illustration of the sample preparation workfl ow. The samples were analyzed by reversed-phase LC-MS. b) Deconvoluted mass spectrum of the Fc/2 subunit. c) Deconvoluted mass spectrum of the GLP-1 peptide. d) Relative amount of the identifi ed Fc/2 variants, displaying the mean value between the triplicate digests and error bars representing the standard deviation. The digestion products were separated by reversed-phase chromatography (BioResolve™ RP mAb Polyphenyl, 450 Å, 2.7 μm 2.1 × 100 mm, Waters™) and analyzed with ESI-QTOF MS (Bruker Impact II). e) Schematic image of the fl exible linker, connecting the GLP-1 peptide to the Fc/2 subunit, and the identifi ed digestion sites.