Product Type
Kit
Description
The complete kit also contains isotopically labeled three most common α-2,3-sialylated glycan standards. Although they are naturally present in small amounts in the Fc portion of antibodies, they can strongly affect effector functions.
Applications
The kits, are low- and high-throughput glycan analysis kits, based on highly pure synthetic isotope-labeled internal standards for the absolute quantification of mAb glycosylation by MALDI-TOF MS. Minimal handling and time are required for sample preparation and within seconds, all relevant glycans of a deglycosylated mAb sample are quantified by comparison of MALDI profiles with the corresponding internal reference standards.
Principle
The glycan analysis is based on the isotopic dilution method and applied to the absolute determination of N-glycans in monoclonal antibodies by MALDI-TOF MS. Isotope dilution analysis is a method of internal standardization, based on the dilution of a mixture of analytes with their isotopically-enriched equivalents, allowing analysis by means of signal ratios. Isotopic dilution is considered a reference method for the absolute quantification of analytes in a mixture.
Size
1 kit
Detection Method
Colorimetric/Fluorometric
Sample
Cell culture supernatant, Urine, Serum, Plasma, Tissue, Adherent cells, Suspension cells, Tissue Homogenate
Protocol Length
1h 00m
Components
Filter device (10 x Amicon 30K spin filter)
Component A. PNGase F (lyophilized)
Component B. standards (Complete Kit 1.6, 100 µL)
Component C. Stabilization reagent (10 mg)
Component D. Sodium citrate tribasic (1 mM solution, 1 mL)
Component E. MALDI-TOF Matrix (DHB, 2 mg)
SDS (2% solution)
Ultra pure water (15 mL for mAb desalting)
USB flash drive. Glycan Quantification Software
Two empty bottles, labeled as HPLC grade Acetonitrile and HPLC
grade Ethanol to be filled by the user.
Requires
Table-top shaker with heating unit.
Centrifuge suitable for 2 mL Eppendorf tubes (floor or table-top).
Centrifugal evaporator suitable for 2 mL Eppendorf tubes.
HPLC grade acetonitrile (1 mL). Do not use lower purity.
Acetic acid (1 mL).
Storage
Please refer to protocols.
Shelf Life
12 months
Research Use
For research use only. Not for human or drug use
