GlycOCATCH™ Smart Enzymes

GlycOCATCH is an enrichment resin for affinity purification of mucin-type O-glycosylated proteins and peptides.

Product Description

GlycOCATCH is designed to specifically bind mucin-type O-glycosylated proteins and peptides. The affinity resin is based on inactive OpeRATOR® enzyme that has been engineered to bind O-glycosylated proteins and peptides with high affinity. O-glycoproteins and peptides are bound to the resin at pH 5-8.

 

GlycOCATCH is provided in a spin column format to allow easy-to-use enrichment of mucin-type O-glycoproteins. The binding is performed in physiological buffers at pH 5-8 and combined with desialylation using SialEXO for 30 min. Due to the strong interaction between the GlycOCATCH resin and O-glycoproteins/peptides, the elution is performed with 8 M urea. Alternatively, the elution can be performed with the included, active OpeRATOR enzyme, that digests the bound O-glycoproteins N-terminally of the O-glycan sites. In this case, peptides are recovered under native conditions. Since asialylated O-glycans on O-glycoproteins are bound efficiently to the resin, the sialidase mix SialEXO® is included in the box.

 

Binding Capacity and Unit Definitions

50 μl of the GlycOCATCH resin binds ≥ 50 μg asialylated etanercept when incubated in 10 mM sodium phosphate buffer pH 7.4, 150 mM NaCl at room temperature for 30 min.

 

One unit of SialEXO hydrolyzes sialic acids from ≥ 90% 1 µg glycoprotein (fetuin) when incubated in 20 mM Tris pH 6.8 at 37°C for 2h.

 

One unit of OpeRATOR digests ≥ 90% of 1 µg glycoprotein (TNFαR) when incubated together with one unit of SialEXO in 20 mM Tris pH 6.8 at 37°C for 2 h.

 

Content and Storage

The GlycOCATCH box (G3-OC6-002) includes:

4 Microspin columns containing GlycOCATCH resin supplied in 20% EtOH, with no preservatives added. Each column includes sufficient material to bind 50 µg O-glycoprotein

1 vial of SialEXO (200 units) supplied lyophilized in TBS pH 7.6, with no preservatives added.

1 vial of OpeRATOR (200 units) supplied lyophilized in TBS pH 7.6, with no preservatives added.

 

The GlycOCATCH box is shipped cold. The GlycOCATCH resin should be stored at +4-8 °C upon arrival. Do not freeze the columns! The vials of SialEXO and OpeRATOR should be stored at –20 °C upon arrival. After reconstitution, SialEXO and OpeRATOR are stable for 1 month at +4-8 °C.

 G3-OC6-002

The GlycOCATCH resin is provided in a convenient spin column format. The box contains 4 GlycOCATCH Microspin columns with a capacity to bind 50 µg mucin-type O-glycoprotein each (200 µg total). GlycOCATCH is provided together with 200 units of SialEXO®, for removal of sialic acids, and 200 units of OpeRATOR®, an optional elution method of the bound O-glycoprotein.

 

Key Characteristics:

Binds mucin-type O-glycosylated proteins and peptides

2h binding

Desialylation using SialEXO® (included) increases binding

Glycoproteins and peptides carrying mucin-type O-glycans

 

 

For more information about GlycOCATCH™ Smart Enzymes