S Cartridges

Specifications for S Cartridges

Application:  For purification of glycans from a variety of complex mixtures including post-labeling cleanup of fluorophore labeled glycans.

Description:  The cartridges contain a glycan binding membrane. This binds to glycans in solutions containing high levels of certain organic solvents (e.g. acetonitrile). Most hydrophobic non-glycan contaminants (e.g. aromatic dyes and detergents) either simply pass through the cartridges or bind very lightly and can be washed off the membrane. The glycans are eluted from the membrane with water.

Binding capacity: Each cartridge can bind up to 20 µg of glycan in a volume no greater than 15 µL.

Number of Samples: S cartridges are designed for single use only.

Suitable Samples: A wide range of glycans can be purified. These include N-linked and O-linked type oligosaccharides, tri-saccharides and larger structures. Substitute 96% acetonitrile with 100% acetonitrile for O-glycan clean-up methods.

Storage: Store at room temperature in the dark. Protect from sources of heat, light, and moisture. The cartridges are stable for at least two years as supplied.

Shipping: The product can be shipped at ambient temperature.

Handling: Ensure that any glass, plasticware or solvents used are free of glycosidases and environmental carbohydrates. Use powder-free gloves for all sample handling procedures and avoid contamination with environmental carbohydrate.

Safety: Please read the Material Safety Data Sheets (MSDS's) for all chemicals used. 

All processes involving hazardous reagents should be performed using appropriate personal safety protection - eyeglasses, chemically resistant gloves (e.g. nitrile), etc. - and where appropriate in a laboratory fume cupboard.

For research use only. Not for human or drug use

Additional Reagents and Equipment Required

• Pure water, HPLC grade (~ 3 ml per sample)

•  Acetonitrile, HPLC grade (~ 3 ml per sample)

• 96 % acetonitrile* / 4 % water (v/v) (~ 6 ml per sample) 100% acetonitrile for O-glycan cleanup.

• 30 % acetic acid / 70 % water (v/v) (~ 6 ml per sample)

Introduction

S cartridges have been designed for use after fluorescent labeling of glycans for the purification of the fluorophore labeled glycans from excess 2AB/2AA dye.

Time Line for Cleanup

S glycan cleanup procedure typically takes around 80 - 100 minutes :

Procedure	Time	Elapsed Time (minutes)
Wash and prime cartridges	20min	20
Apply sample	20min	40
Wash off non-glycan contaminants	20min	60
Elute glycans	20min	80

Instructions for Use

1）Prepare the glycan sample

The sample to be cleaned must have a volume of 15 µl or less. If your sample has a greater volume then dry it down using a centrifugal evaporator and reconstitute in not more than 15 µl of water.

2 ）Prime the S cartridge

Prime the S cartridges (one per sample) by washing each with the following :

• 1st wash - 1 ml water.

• 2nd wash - 5 ml 30 % acetic acid (aq). 

• 3rd wash - 1 ml acetonitrile.

Allow each wash to drain completely before adding the next. If flow is restricted, e.g. by an air gap, apply a slight pressure to the top of the cartridge in order to resume normal flow.

3）Spot sample onto cartridge membrane

Spot each sample onto a freshly washed cartridge disc ensuring that the disc is still wet with acetonitrile.

Spread the spot over the entire disc surface if possible as this aids cleanup.

4）Allow sample to adsorb onto membrane

Allow adsorption for 15 minutes.

5） Add residual sample from sample vial

Rinse each sample vial with 100 µl acetonitrile and apply to the corresponding cartridge disc.

6）Wash non-glycan contaminants off membrane

Wash each disc with 1 ml acetonitrile, followed by 5 x 1 ml 96 % acetonitrile* / 4% water 

*(for cleanup of O-glycans or N- and O- glycans labeled with procainamide, substitute with 100% 

acetonitrile)

7）Elute glycans off membrane into a suitable container by eluting with 2 x 0.5 ml water

Allow each 0.5 ml aliquot to drain before the next is applied.

8）Dry the eluted glycans (optional)

If appropriate, evaporate the glycan containing fraction to dryness, then dissolve in a desired volume ofwater or solvent for further analysis.

9）Protocol Complete

Your glycans are now ready to analyse.