Glycan Profiling

No. Lectenz® are lectin-like, enzyme-derived binding proteins derived from inactivated glyco enzymes and optimized through directed evolution. This results in a recombinant binding reagent that has high specificity, unlike some lectins, and is monomeric.

Absolutely. Email sales@lectenz.com or call +1 (706) 549-4484 between 9AM-5PM ET and we’ll be happy to assist you.

Our product quality is ensured through a rigid Quality Management System that combines documentation assurance and rigid analytics. For more information, see our Quality Commitment.

The GlycoSense™ array is similar to a lectin array, except that in GlycoSense™, lectins (or other glycan-binding proteins) are conjugated to 6 μm microspheres instead of a flat array surface.  This allows each lectin-conjugated microsphere to be counted on a flow cytometer, enhancing statistical reproducibility.  The microspheres are spectrally distinct, allowing different glycan-binding microspheres to be combined in a single glycoprotein sample.

GlycoSense™ analysis can be performed with any basic two-color flow cytometer, with a 633/640 nm red excitation laser to detect each microsphere and a 488 nm blue excitation to detect green fluorescently labeled glycoproteins that are bound to the microsphere.

Glycoproteins can be directly labeled using NHS esters of 488 excitable green fluorescence dyes (Alexa Fluor 488®, DyLight® 488, FITC, SureLight™ 488, etc.), with non-conjugated dye removed using dye removal columns or dialysis.

Yes, instead of direct labeling, a green fluorescent secondary binding molecule, such as a binding partner or a Fab fragment against the glycoprotein of interest, may be used to indirectly label the glycoprotein analyte. The use of full antibodies as a secondary label could be problematic, however, as antibodies themselves are glycosylated.

Antibody glycosylation can be difficult to detect if the glycan is buried inside the core Fc of antibodies. While we have been able to monitor enzymatic modification on the glycans of some antibodies, we have had difficulty with others. Lectenz Bio can run preliminary assays for an antibody you are interested in monitoring using GlycoSense™.

GlycoSense™ data are statistically reproducible and repeatable, but due to differences in the inherent affinities of each GlycoSense™ capture reagent and the dependence of affinity on glycan structure, absolute glycan quantitation is not possible using GlycoSense™.  However, GlycoSense™ signals between replicates or related samples can be compared to determine relative change in glycan feature i.e., to determine whether two batches of a protein sample have similar glycan profiles, or whether a protein sample has less of a specific glycan feature after glycosidase treatment.

No, a GlycoSense™ analysis only reports total signals for specific glycan features (terminal Sia, Gal, GlcNAc, etc., in some cases with associated glycosidic linkage information) present on a glycoprotein.  A GlycoSense™ analysis allows you to evaluate changes in glycosylation features in glycans.  This platform provides data that is complementary to traditional glycoprofiling.

GlycoSense™ can be performed rapidly, typically in as short as one hour using a basic two-color flow cytometer and does not require the enzymatic or chemical release of glycans from the protein, sometimes without pre-purification of the glycoprotein from media or buffer.  The simplicity of the process greatly reduces the level of personnel training and does not require highly specialized instrumentation, making it convenient for adoption by individual research groups, industrial settings, and core facilities.  Rather than providing a list of observed glycans, GlycoSense™ analysis provides an aggregate “snapshot” of glycan features, which can be particularly convenient for monitoring in-process biologics production or in vitro glyco enzyme reactions (glycoengineering).

GlycoSense™ can be used to analyze the differences in glycan features between glycoprotein samples.  This can be applied to monitor changes in the glycosylation features during the production of glycoprotein biologics in near real-time, or to compare the glycosylation features of glycoprotein production lots, or to monitor the progress of glycoengineering of a glycoprotein. It can also easily provide linkage information; comparing α2,3 versus α2,6 sialic acid linkages between samples is particularly convenient with GlycoSense™.

Yes, we can manufacture customized GlycoSense™ kits with glycan-specific microspheres to suit your applications.  For questions about custom applications, please contact us via our web form or email info@lectenz.com.

Yes. Lectenz Bio scientists are available to help you solve any technical issue you may encounter with the platform and its adaptation to your custom applications. Contact us via our web form or email info@lectenz.com with any questions.