PNGase F PRIME™ - Especially Optimized for Mass Spec Imaging
Matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS) is a unique and well developed tool for analyzing the molecular content of frozen or formalin-fixed, paraffin embedded tissue (FFPE). PNGase F PRIME™ is the only commercially available version of Peptide-N-Glycosidase F that is validated and optimized for use in imaging mass spectrometry.
Within the past two decades, MALDI Imaging has shown substantial potential for clinical applications. Studies have tested and proven the instrumentation’s applicability to different clinical challenges such as tumor staging, receptor identification, drug delivery and drug efficacy. Additionally, an approach utilizing this instrumentation for the analysis of glycosylation in formalin-fixed paraffin embedded (FFPE) tissue has gained monumental traction in the field.
The key to this approach is the spraying of a molecular coating of peptide N-glycosidase onto the tissue to specifically release N-glycans. This method not only allows for the discovery of N-linked glycosylation changes within a tissue, but it also allows for these changes to be spatially localized and correlated to the tissue’s histopathology. Because of the method’s applicability to FFPE tissues, any pathology block prepared by standard workflows has the potential to be analyzed. This approach has allowed for the assessment of N-linked glycans in xenograft tissues, FFPE clinical tissues and tissue microarrays.
Since the introduction of this method, the concept of spraying an enzyme onto a surface for MALDI Imaging analysis has been applied to a variety of other applications, including cell culture, whole serum/plasma analysis, and glycoprotein capture through antibody arrays. Using this principal methodology and interchanging enzymes and/or biological material of interest exponentially elevates the possibilities of scientific research and discovery.
While the number of glycosylation studies utilizing MALDI Mass Spectrometry Imaging (MSI) is rapidly increasing, previously MALDI MSI was mainly utilized for analyzing peptides and lipids. In 2014, Powers et. al. published the first study using MALDI MSI to profile N-glycans in formalin-fixed paraffin embedded (FFPE) tissue blocks and tumor microarrays.
Since this study, many others have employed the methods described to analyze glycosylation changes in various disease states. In 2018, Angel et. al. published the first study to analyze N-glycans and tryptic peptides on the same FFPE tissue section using MALDI MSI, allowing for the identification of a colocalized protein population linked to the exact same regions of N-glycans. In 2020, McDowell et. al. published a study that combined MADLI MSI and lectin analysis of N-linked glycans in carbohydrate antigen-defined pancreatic cancer tissues and found distinct differences not only in N-glycan spatial localization across both healthy and diseased tissues, but also between different biomarker-categorized tissues.
In addition to the multitude of MSI studies analyzing glycosylation changes in tissues, new methods have emerged using MSI for analysis of glycans on antibody-captured proteins, whole serum and plasma, and cultured cells. In all of these studies, N-Zyme Scientific’s products have been optimized and applied to ensure the best results possible.