Waters GlycoWorks Single Use Sample Preparation Kit - Manual

GlycoWorks High-throughput Sample Preparation Kit 1
[ CARE AND USE MANUAL ]
LC-FLR
LC-FLR-MS
I. INTRODUCTION
This document provides information regarding the general care and
use of the Waters GlycoWorks™ Single Use Sample Preparation Kit
for release of N-glycans. This protocol can be used as a guideline and
is validated using IgG but can also be used and optimized for other
N-linked glycoproteins. This sample preparation offers an inclusive
protocol that allows the user to source many of the critical components
needed to perform the preparation from one vendor.
The Single Use Catridge Pack contains 20 1cc catridges, good for
10 analyses.
For comparison results using this protocol versus the 96 well plate
and for recovery information with the control standard
1,2 .
The guide provides the following:
■
An instruction template for new users that highlights the use of a
control standard (included in the kit) for first time validation as
well as subsequent troubleshooting
■
Protocol for LC/FLR and LC/FLR/MS
■
Tips and tricks throughout each section to help aid in the success
■
An online training video is also available 3
GlycoWorks Single Use Sample Preparation Kit
CONTENTS
I. INTRODUCTION
II. STORAGE AND DISPOSAL OF CARTRIDGES
III. ABBREVIATIONS
IV. USING THE GLYCOWORKS SINGLE USE KIT
V. PREPARING LABELED GLYCANS FOR HILIC-FLR
WITH AN ACQUITY UPLC BEH GLYCAN COLUMN
VI. REFERENCES
VII. ORDERING INFORMATION
Figure 1: Sample Preparation from Glycoprotein to Labeled Glycans
Glycoproteins
FLR Labeling Reaction of Glycans
3
FLR Labeled Glycans
MALDI MS
Denaturation and Deglycosylation
RapiGest™ (if necessary)
DTT Reduction (if necessary),
IAM Alkylation (if necessary)
Deglycosylation by PNGase F
1
Extraction of Released Glycans
GlycoWorks HILIC 1 cc Cartridge
2
Excess Labeling Reagent Removal
GlycoWorks HILIC 1 cc Cartridge
4
General Guideline of Sample Preparation from Glycoprotein to
enrich FLR Labeled Glycans Using Reductive Amination Reaction.
If using MALDI please refer to Product Solution, Literature Code 720004660EN