SmartEnzymes GlycOCATCH workflow

Enrichment of O-glycopeptides

GlycOCATCH is an affinity resin for purification of 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.

The applications of GlycOCATCH include specific enrichment or removal of O glycoproteins and peptides, glycomics, studies of complex samples and characterization of biopharmaceuticals.
 

  • Specific enrichment of mucin-type O-glycosylated proteins and peptides
  • Reliable - high affinity binding
  • Fast and easy-to-use workflow
Mucin-type O-glycosylated proteins and peptides
Binds to glycoproteins and peptides carrying mucin-type O-glycans
2 h workflow
Desialylation using SialEXO® (included) increases the binding

GlycOCATCH® Affinity Purification

Enrichment resin for affinity purification of mucin-type O-glycoproteins and peptides

About GlycOCATCH®

GlycOCATCH workflow

GlycOCATCH is designed to specifically bind mucin-type O-glycosylated proteins and peptides. The affinity resin is based on the inactive OpeRATOR enzyme, that has been engineered to bind O-glycosylated proteins and peptides with high affinity.

The binding is performed in physiological buffers at pH 5-8. Due to the strong interaction between the GlycOCATCH resin and O-glycoproteins/peptides, the elution can be performed with 8 M urea. Alternatively, the elution can be performed with the active OpeRATOR enzyme, that digests the bound O-glycoproteins N-terminally of the O-glycan sites. In the latter case, peptides are recovered under native conditions. The OpeRATOR Lyophilized enzyme is included in the purchase. Since asialylated O-glycans on O-glycoproteins are bound more efficiently to the resin, the sialidase mix SialEXO Lyophilized is also included in the purchase.

GlycOCATCH® Affinity Purification

Enrichment resin for affinity purification of mucin-type O-glycoproteins and peptides.

GlycOCATCH Affinity Purification is provided in a convenient spin column format. The product contains:

  • 4 GlycOCATCH Affinity Purification Microspin columns, each with a capacity to bind 50 µg mucin-type O-glycoprotein (200 µg in total)
  • SialEXO Lyophilized 200 units, for removal of sialic acids
  • OpeRATOR Lyophilized 200 units, for an optional elution method of the bound O-glycoprotein

 

  • Targeted O-glycoprotein enrichment
  • Specific clean-up of complex samples
  • Sample preparation for improved MS analysis

Available Products

GlycoCATCH Affinity Purification Microspin 0.2 mgEnrichment resin for affinity purification of 0.2 mg mucin-type O-glycoproteins and peptidesG3-OC6-002806.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

Unit Definition

  • 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

GlycOCATCH Affinity Purification 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 Lyophilized (200 units) supplied in TBS pH 7.6, with no preservatives added.
  • 1 vial of OpeRATOR Lyophilized (200 units) supplied in TBS pH 7.6, with no preservatives added.

GlycOCATCH Affinity Purification is shipped cold. Content should be stored at different temperatures upon arrival!
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.

FAQ and Support

Popular FAQ

The OpeRATOR enzyme digests O-glycosylated proteins N-terminally to the serine (S) and threonine (T) glycosylation sites. OpeRATOR therefore facilitates mapping of O-glycosylation sites by LC-MS and as the digested products no longer bind to the GlycOCATCH resin, elution and sample preparation can be achieved in a single step. However, this requires the O-glycosylation sites to be spaced closely together for the OpeRATOR digestion to yield peptides of a size suitable for analysis by LC-MS/MS.

Yes. IMPORTANT! Before you load your sample make sure that the protease in solution is inactivated and that the pH, buffer, chaotrope and NaCl concentrations are within the recommended ranges according to the instructions.

No, GlycOCATCH is designed to bind specifically to mucin type (core 1) O-glycosylated proteins and peptides. The performance is significantly enhanced by removal of sialic acids using SialEXO.

No, when the glycoprotein is digested with OpeRATOR, the binding capacity of GlycOCATCH is lost.

 

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