N-Glycan Kit (2AB-Magnetic Beads)

Product Overview

Typical characterization of N-glycan profiles involves cleaving glycan chains with PNGase F, labeling with the derivatization reagent of 2-AB, followed by analysis via high-performance liquid chromatography (HPLC). For the conventional 2-AB labeling workflow, enzymatic digestion duration and derivatization labeling time are the two primary rate-limiting steps. This N-glycan analysis kit of 2-AB (magnetic bead method) enables rapid enzymatic digestion, labeling and enrichment, with the entire sample pretreatment procedure taking less than 60 minutes.

Key Features

Simplified workflow

Integrates enzymatic digestion, 2-AB labeling, and enrichment/purification into a streamlined workflow consisting of three main steps.

Rapid enzymatic digestion

Enzymatic deglycosylation can be completed in approximately 10 minutes at 50 °C under recommended conditions.

Fast labeling reaction

2-AB derivatization labeling can be completed in approximately 15 minutes at 65 °C.

Magnetic bead-based enrichment and purification

Magnetic bead separation eliminates the need for lyophilization steps. Typical recovery rate is >90%, with a processing time of ≤15 minutes per sample.

Improved sample handling compatibility

Centrifugal filter tubes provided in the kit can be used at 8,000 × g for 5 minutes to reduce potential interference from residual magnetic beads in downstream chromatographic analysis.

Applications

Rapid N-glycan profiling

Designed for rapid preparation and analysis of N-linked glycans released from glycoproteins for downstream HPLC-based characterization.

Streamlined glycan sample preparation workflow

Supports efficient sample pretreatment for laboratories performing high-throughput or time-sensitive N-glycan analysis.

Compatibility with 2-AB labeling-based analytical methods

Suitable for workflows based on 2-AB derivatization for glycan detection and profiling in research applications.

Performance Data

Performance Summary

No.	Performance	Test Protocol	Criteria		Results of Hzymes
1	Blank Control	1. Prepare IgG samples and blank solution according to the IFU	No interfering peaks in blank solutions		YES
1	Blank Control	2. No interfering peak shall appear at the retention time of the peak in IgG sample for the blank solution	No interfering peaks in blank solutions		YES
2	Repeatability	1. Prepare 6 parallel samples of IgG	G0F	Area CV ≤ 15%	0.23%/2.43%/1.17%
2	Repeatability	2. Calculate the CV of G0F area &G0F% of the 6 samples	G0F	%Area CV ≤ 10%	0.11%/1.86%/1.74%
3	Intermediate Precision	1. Prepare 6 parallel samples of IgG by two persons separately	G0F	Area CV ≤ 15%	4.53%/7.62%/5.13%
3	Intermediate Precision	2. Calculate the CV of G0F area &G0F% of the 12 samples	G0F	%Area CV ≤ 10%	1.00%/2.29%/1.64%
4	Inter-Batch Variation	1. Take 3 batches of reagents, and prepare 3 samples with 3 replicates per batch independently	G0F	Area CV ≤ 15%	6.02%
4	Inter-Batch Variation	2. Calculate the CV of G0F area &G0F% between batches respectively	G0F	%Area CV ≤ 10%	2.33%
5	Linearity	1. Prepare linear solutions of IgG/Etanercept (at least 5 points from 30 μg to 180 μg)	R² of Area and concentration of G0F:R²≥0.990		0.9985/0.9938
5	Linearity	2. Calculate the correlation coefficient R² of sample concentration and G0F peak area	R² of Area and concentration of G0F:R²≥0.990		0.9985/0.9938

Inter-Batch Variation

G0F	Area			% Area
Batch Number	Lot1	Lot2	Lot3	Lot1	Lot2	Lot3
1	514085285	514457578	474229594	21.4	21.45	22.36
2	512370466	491897780	483163068	21.34	21.59	21.43
3	513192769	486313705	480701026	21.35	22.03	21.44
4	514417080	493153056	479721706	21.35	22.41	21.33
5	515701835	512146723	490068853	21.36	21.37	21.52
6	514603122	489866450	476050234	21.38	22.01	21.6
Mean	514061760	497972549	480655747	21.36	21.81	21.61
SD	1159771	12119663	5626258	0.02	0.4	0.38
CV	0.23%	2.43%	1.17%	0.11%	1.86%	1.74%
CV requirement 1	CV≤10%			CV≤10%
Mean (total)	497563352			21.6
S2 (total)	897310036558175			0.253
CV (Inter-Batch)	5.82%			1.80%
CV (total)	6.02%			2.33%
CV requirement 2	CV≤15%			CV≤10%

Linearity

FAQ

How to deal with residual magnetic beads in samples after processing?

Magnetic beads may remain in the sample after purification. It is recommended to perform a centrifugation step using the matched filter tube prior to instrumental analysis (8000 × g for 5 min, or adjust time appropriately depending on residual liquid volume).

For high-throughput sample processing, centrifugation time may be extended to more than 10 minutes to ensure more complete bead removal.

How should samples be handled before instrumental analysis to ensure stable results?

Since the final sample volume after dilution is relatively small and the buffer contains volatile acetonitrile, it is recommended to minimize evaporation effects that may impact peak area and peak shape.

Best practice includes:

Injecting samples into the instrument immediately after centrifugation, or

Transferring samples to EP tubes and storing them in a sealed state (recommended storage time ≤ 3 days)

Avoid prolonged storage in open sample vials before injection

Are there any additional recommendations for high-throughput analysis?

For workflows involving a large number of samples, it is recommended to install a protective guard column (pre-column) to reduce contamination risk and extend the service life of the analytical column.