PATH® Protein Microarrays combined with MALDI MS for high throughput N-Glycan analysis.

IN PATH non-porous nitrocellulose

Emerging studies in the field of glycobiology are beginning to reveal a complex and diverse realm of critical and abundant sugar-based polymers that support a wide range of biological processes.   Glycans cannot be studied by molecular weight alone and they have an exceptionally diverse range of structures and functions, making them difficult to study.  The US National Institutes of Health established the Common Fund Glycoscience, seeking to further this understanding.  In this article, Alyson P. Black, et al describe a pioneering method combining MALDI-MAS spectrometry imaging with nitrocellulose planar antibody arrays. Antibodies are applied to Grace Bio-Labs PATH® protein microarrays, blocked, and serum samples applied using standard methods.  Here, the thin-film nitrocellulose PATH® slides provide a stable and high capacity binding substrate for the proteins, while the nitrocellulose layer remains thin enough to be compatible with mass spectrometry applications.  Glycans are then enzymatically cleaved using a spraying method retaining glycan co-localized array positions relative to the antibodies.  MALDI MS was used to image N-glycans for location and intensity using standard methods.  Results were confirmed by HPLC for intensity and lectin binding for locations.  This method provides a novel tool where 100’s or 1,000’s of glycans can be analyzed in a single sample imaging run, providing sensitivity, specificity, and throughput greater than ELISA.  


Novel Mass Spectrometry Platform for Multiplexed N‑Glycoprotein Biomarker Discovery from Patient Biofluids by Antibody Panel Based N‑Glycan Imaging, Alyson P. Black, et al.