EZGlyco® mAb-N Kit with APTS | Sumitomo Bakelite Co., Ltd.

Products

S-BIO confidently offers an antibody (IgG) N-glycan sample preparation kit compatible with capillary electrophoresis (CE). This kit allows to start from purified IgG or unpurified IgG (such as the culture supernatant of CHO cells), and perform IgG immobilization, N-glycan release, fluorescence labeling and clean-up all in one tube, enabling quick and simple preparation of glycan samples for CE analysis. The kit includes a glycan release enzyme and a labeling reagent (APTS). Moreover, the labeling reaction does not use hazardous cyanide compounds, making it a user-friendly kit.

Features

Significantly shortened sample preparation time

Complete glycan sample prep in 4 hours including IgG purification process

Universal fluorescent label for CE

Compatible with APTS labeling, widely used in CE glycan analysis
Non-toxic reducing agent used in the labeling process

All essential reagents included

N-Glycosidase, labeling reagents and other necessary reagents are included

Application

High-throughput Analysis of Glycans in Biologics Research and Development

Supports high-throughput glycan analysis by CE in manufacturing process development
Glycan analysis using a 1st generation DNA sequencer (capillary electrophoresis type)

Example of Analytical Conditions

[CE instrument]

P/ACE MDQ Plus (AB Sciex)

[Analytical conditions]

Capillary: N-CHO capillary (AB Sciex, 477601)
Buffer: N-Linked Carbohydrate Separation Gel Buffer (AB Sciex, 477623)
Sequence: Shown in the table
Fluorescent detection: Ex. 488 nm, Em. 520 nm

Table: Analysis sequence

EZGlyco® mAb-N Kit with APTS Protocol (665KB)

Example of Analysis Data

N-glycan analysis of IgG in cell culture supernatant

A mixture of human serum IgG (40 μg) in the culture supernatant (120 μL) of HEK293 cells was treated with the EZGlyco® mAb-N Kit with APTS to prepare APTS-labeled N-glycan samples. Purified human serum IgG was treated in the same way as a control. CE analysis was performed under the conditions described.

The same peak pattern was obtained with the IgG spiked in the culture supernatant and the purified IgG, demonstrating that this kit can be applied to both purified and unpurified antibodies.

N-glycan analysis of Antibody-Drug Conjugates (ADC)

(a) Trastuzumab deruxtecan and (b) Trastuzumab emtansine (40 μg) were treated with the EZGlyco® mAb-N Kit with APTS to prepare APTS-labeled N-glycan samples. CE analysis was performed under the conditions described.

Data demonstrated that this kit can be applied to not only naked IgG but also ADC N-glycan analysis.

Specifications

Product No.	Product Name	Contents	Quantity	Price
BS-43607Z	Auto-EZGlyco® mAb-N Kit with APTS	Antibody capturing column Antibody capturing buffer Washing buffer PNGase F PNGase F enhancer APTS (8-aminopyrene-1,3,6-trisulfonic acid) Reducing reagent (non-toxic) Cleanup column Protocol booklet	10 tests	ASK