The protocol used for the implementation of the GlycoWorks RFMS kit on Andrew+ is the quality control (QC)/automation-friendly sample preparation method. The QC GlycoWorks RFMS protocol was validated by Waters on Andrew+. Experimental conditions and prameters were fine-tuned to fit the hardware configuration of the Andrew+ robot, i.e., labware Dominos and required, additional, connected devices, to produce reliable results comparable to target scores. The QC protocol is an adaptation of the standard variable volume (VV) method in which the starting sample concentration and reagent concentrations used during protein denaturation, deglycosylation, and N-glycan labeling have been carefully modifed to allow pipetting volumes of 10 uL or more thus improving pipetting accuracy and generating results comparable to those produced by the standard VV method. GlycoWorks RapiFluor-MS N-Glycan Eco Kits replace DMF with a greener, safer, alternative (DMSO), as a Reagent Solvent for the RapiFluor-MS Reagent Label and remove any other DMF containing solutions. The kit includes the following: the GlycoWorks Deglycosylation Module, GlycoWorks Eco Labeling Module, GlycoWorks Eco Clean-up Module, and GlycoWorks SPE Reagents for Automation. It contains materials to process 96 samples in a 2x48 format. The kit contains: (2) 0.08 ml/vial Rapid PNGaseF Enzyme (1) 1.5 ml/vial Glycoworks Rapid Buffer (3) vials of 20 mg/vial RapiGest SF Surfactant (1) Intact mAb Mass Check Standard *Shipped in a separate box; GlycoWorks RapiFluor-MS Eco Labeling Module: (2) 55 mg/vial RapiFluor-MS Reagent (2) 1 ml/vial Reagent Solvent (Anhydrous DMSO); GlycoWorks RFMS Eco Clean-up: (1) GlycoWorks HILIC µElution Plate; GlycoWorks SPE Reagents - Automation: (2) 12 mL/vial Elution Buffer, (2) 18 mL/vial Equilibration Solvent; (2) 70 mL/vial Wash Solvent
Suggested Use: Eco-Friendly, Released N-linked Glycan Analysis, Andrew+ Automation
