Innovative Biochemical Analysis Solutions

Typical Binding Assay Result from the Mucin Glycan Array

Mucin Glycan Array Features

Mucin Glycans and Cancers

Example 2:  Mucin glycan array on a Z Biotech’s NHS slide compared with a competitor slide. Subarrays were assayed with glycan-binding protein biotinylated Peanut agglutinin (PNA) lectin (10 μg/mL), followed by Streptavidin-Cy5.  Array was scanned with GenePix scanner at 545 PMT and 100% laser power at 635nm wavelength.  There is very low non-specific binding for the negative control spots.  Positive control 1 and the marker show binding as expected, as well as β1,3-galactose-containing O-glycans (T antigens and Mucin O-GalNAc Core 2). Analysis using Genepix software produced data to allow a comparison of background signal, relative fluorescence units (RFU), and signal-to-noise ratio (SNR) between slides.

Mucin Glycan Array

Autoantibodies to cancer-specific antigens have the potential to be biomarkers for early detection of cancer as well as cancer immunotherapeutic agents. Extracellular mucins such as MUC1 are overexpressed and aberrantly glycosylated in many cancers. The extracellular domain of the MUC-1 contains a variable number of tandem repeats (VNTR) of 20 amino acid residues with serine (Ser) or threonine (Thr) sites for O-glycosylation. A recent study indicated that high levels of a subset of autoantibodies with an immunodominant epitope of GalNAca1-O-Ser/Thr (Tn antigen) and Neu5Aca2-6GalNAca1-O-Ser/Thr (STn antigen) on MUC1 were significantly associated with reduced incidence and increased time to metastasis of breast cancer. Thus the Mucin Glycan Array can serve as a research tool for determining mucin glycan immunodominant epitopes in cancers and furthermore sensitive detection of expression of autoantibodies.

Example 1:  Mucin glycan array on a Z Biotech’s NHS slide compared with a competitor slide. Subarrays were assayed with glycan-binding protein biotinylated Helix pomatia agglutinin (HPA) lectin (10 μg/mL), followed by Streptavidin-Cy5.  Array was scanned with GenePix scanner at 545 PMT and 100% laser power at 635nm wavelength.  There is very low non-specific binding for the negative control spots.  Positive control 1 and the marker show binding as expected, as well as terminal-GalNAc-containing O-glycans (Tn antigens, MUC-1 glycopeptides, T antigens). Analysis using GenePix software produced data to allow a comparison of background signal, relative fluorescence units (RFU), and signal-to-noise ratio (SNR) between slides.

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