Presentation

Ludger V-Tag Glycopeptide Labeling and Enrichment Kit

We are pleased to announce the launch of a kit for the labeling and enrichment of IgG glycopeptides, enabling analysis by (U)HPLC or MALDI Mass Spectrometry.

The V-Tag kit (Cat # LT-VTAG-24) is suitable for IgG subclass glycoproteins that have been digested with pronase or trypsin to release the glycopeptides.
As little as 5ug IgG sample can be used. The V-Tag system is comprised of two steps which can be completed in 2 hours. The first step involves the labeling of peptides and glycopeptides from the protease digest using a novel fluorophore (1 hour). The second step is the enrichment of the glycopeptides using a solid phase extraction (SPE) hydrophilic interaction liquid chromatography (HILIC) cartridge (1 hour).

This kit has been validated according to ICH guideline Q2 (R1) guidelines. Using different IgG samples and replicates of 9 for each, CVs for repeatability were typically <5%.

Presentation Quick Links:

1. Introduction: mAb Glycopeptide Profiling with V-Tag
2. Highlights of the V-Tag System
3. Studies using V-Tag at Ludger
4. Incorporating V-Tag into your drug programme
5. Next Steps

 Downloadable pdf version of presentation

Poster

A Fluorescent Labeling and Enrichment System for Glycopeptides Generated from Proteolytic Digestion of IgG mAbs; A System That Can Be Used as Part of the Peptide Mapping Workflow

Hendel JL, Badia-Tortosa C, Spencer DI, Fernandes DL
Presented at: USP: Glycosylation Analysis for Biopharmaceuticals Workshop
Washington DC, United States. August 2015

View product documentation

Product guide

Photo procedure

Product specification

Application 
For the fluorophore labeling and enrichment of glycopeptides

Description 
This kit contains the reagents needed for the conjugation of V-tag dye (LT- VTAG-01) to the amine moieties of glycopeptides, and the specialised solid phase extraction (SPE) cartridges required for the purification and enrichment of the labeled glycopeptides.

Dye Properties
Mass = 434.42 (resulting in a glycopeptide mass increase of 319.33 Da.)
Fluorescence, λex = 250 nm, λem = 360 nm.

Number of Samples
24 separate analytical samples per kit.

Amount of Sample
From approximately 1 µg to 50 µg of glycoprotein per sample. 

Suitable Samples
IgG single subclass glycoproteins. Other glycoproteins can be used, but note that if there are multiple glycosylation sites, the glycopeptides may need to be separated by C18 HPLC before HILIC analysis. It is recommended that samples do not have amine containing solvents (e.g. Tris type buffers) or reduction/alkylation agents as they will interfere with the V-tag labeling. Removal of the amine containing solvents and reduction/alkylation agents can be accomplished using devices which are designed for desalting and buffer exchange, as well as removal of low-molecular weight compounds.

Labeling Selectivity
One V-tag label for every IgG glycopeptide N-terminus

Storage: Store the V-tag dye at -20°C in the dark. Protect from sources of heat, light and moisture. Once used, extra dye solution can be frozen and re-used. 

Handling: Ensure that any glass, plastic-ware or solvents are free from unwanted peptidases, glycosidases and environmental carbohydrates. Use powder-free gloves for all sample handling procedures and avoid contamination with environmental carbohydrates.

Safety: For research use only. Not for human or drug use . Please read the Safety Data Sheets (SDS's) for all chemicals used. All processes involving labeling reagents should be performed using appropriate personal safety protection – safety glasses, chemically resistant gloves (e.g. nitrile), lab coat, and when appropriate, in a laboratory fume cupboard.

Each labeling kit consists of each of the following: