Lectenz Bio : Glycan Detection | Processing | Profiling
Lectenz Bio : Glycan Detection | Processing | Profiling

FAQ

FAQ

Science Questions


Are Lectenz® lectins?

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.

I have bought a Lectenz® kit, but I’m having trouble getting my assay to work. Can you help?

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

How do you ensure consistent quality in your products?

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.

How does the GlycoSense™ Array Kit differ from a lectin array?

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. 

What equipment is required to use a GlycoSense™ Array?

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.

How can I directly label my glycoprotein for GlycoSense™?

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.

Can I use a fluorescent secondary reagent to detect my glycoprotein in GlycoSense™?

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.

Can GlycoSense™ be used to detect antibody glycosylation?

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™.

Are the fluorescence signals from GlycoSense™ quantitative?

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.

Will GlycoSense™ tell me the glycoforms on a glycoprotein and their percent abundance?

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.

What are the advantages of GlycoSense™ over traditional MS or HPLC analyses?

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).

What are some of the major uses of the GlycoSense™ platform?

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™.

Are custom GlycoSense™ kits available for specialized applications?

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.

Is technical support available for GlycoSense™ analysis?

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.

Will GlycoSense™ work for me?

If you have specific questions about using GlycoSense™ for your research, please contact us via our web form or email info@lectenz.com. We are more than happy to discuss your research and what the GlycoSense™ platform can do for you.