QA-Bio/Glucose Homopolymer Ladder/CPROC-GHP-30

The glucose homopolymer ladder derived from digested wheat starch used as a standard for sizing of glycans. Amount supplied sufficient for 30 HPLC runs.

View the Glucose Homopolymer Application Note

Sources: Derived from wheat starch

Form: Dry. Dried by centrifugal evaporation from an aqueous solution.

Amount:
CAB-GHP-30 100 picomoles2-AB
CAA-GHP-30 100 picomoles2-AA
CPROC-GHP-30 100 picomolesProcainamide

Purity: > 90% pure as assessed by a combination of HPLC

Storage: -20˚C both before and after dissolution. The products are stable for at least 5 years as supplied. View Certificate of Stabilty

Shipping: The product can be shipped at ambient when dry. After dissolution, ship on dry ice.

Handling: Allow the unopened vial to reach ambient temperature and tap unopened on a solid surface to ensure that most of the lyophilized material is at the bottom of the vial. Gently remove the cap, add the desired volume of reconstitution medium, re-cap and mix thoroughly to bring all the oligosaccharide into solution. For maximal recovery of oligosaccharide, ensure that the cap lining is also rinsed and centrifuge the reconstituted vial briefly before use. Ensure that any glass, plasticware or solvents used are free of glycosidases and environmental carbohydrates. Minimise exposure to elevated temperatures or extremes of pH. High temperatures and low pH will cause desialylation. High pH will cause epimerisation of the reducing terminus GlcNAc.

Safety: The glucose homopolymer ladder is non-hazardous and has been purified from natural sources certified to be free of all hazardous material including pathogenic biological agents.
Download Non-hazardous MSDS

Use of Glucose Homopolymer Ladder as a system suitability standard

GHP ladder can be used as a system suitability standard (See figure 1) to ensure that the liquid chromatography system is fit for purpose. The criteria which must be met are the following:

• Does the GHP profile match the profile shown in the CofA of the standard?
• Are the peaks symmetrical and well resolved? Do the peaks match the regular distribution pattern?
• Do at least two GHP profiles overlay?

Figure 1: Triplicate overlay of Ultra-high performance liquid chromatography (UHPLC) chromatograms from 2-AB labelled GHP Ladder [CAB-GHP-30] run on a HILIC column. Each peak is labelled with their corresponding GU value.

Common problems encountered when assessing system suitability can include the following; asymmetrical peaks may indicate compromised column quality or column aging. Whilst, irregular elution patterns could result from insufficient equilibration and/or various issues with LC hardware components (i.e. pumps, buffer system, blockages etc.)

Use of Glucose Homopolymer Ladder as a reference standard

GHP ladder can also be used as a reference standard (See figure 2) to assign GU values to peaks in the released glycan pool by comparison with the ladder. These GU values are reproducible and predictive as each monosaccharide in a glycan contributes a set increment to the GU value. This allows for primary assignment of structure by comparison of GU values for unknown glycans with glycan standards whose GU values are in databases or reported in the literature.

It is important to remember that the proposed identity of a glycan must be confirmed wi th further orthogonal structural analysis (e.g. MS or exoglycosidase sequencing).

Figure 2: Stack plot of HILIC UHPLC chromatograms from 2-AB labelled GHP (top panel) used as a reference standard for analysis of 2-AB-labelled glycans released from human IgG [GCP-IgG-100U] (bottom panel).