Construction and Analysis of Glycan Microarrays

IN Discussion Topics, ONCYTE nitrocellulose film slides, PATH non-porous nitrocellulose, ProPlate

Protein glycosylation is a complex post-translational modification (PTM) that provides great proteomic diversity. It involves the attachment of sugars at specific amino acid residues. Asn (N-linked) or Ser/Thr (O-linked) being the residues most frequently glycosylated. Glycosylation is critical for physiological and pathological cellular functions and affects the bioavailability, efficacy, stability, and solubility of therapeutic glycoproteins. Therefore, obtaining information about the type, position, and composition of glycans within a protein sequence is paramount. The structural diversity and heterogeneity of the glycoproteins present a significant analytical challenge. Glycan microarrays technology is often adopted for glycosylation studies as it enables high-throughput screening of glycan-mediated interactions.

A recently published review, offer a comprehensive overview of the methodology, including information about:

  • Selection of microarray substrate
  • Printing conditions
  • Assay development
  • Data acquisition and processing.

It also lists tools useful for glycan microarray analysis:

Repositories of glycan microarray data

CFG Database Imperial

College microarray data online portal

CarbArrayART (available for testing upon request)

GlyGen microarray repository

Data visualization/manual data analysis

GLADGLycan Array Dashboard for visualization of microarray data

Automated analysis

MotifFinder: to mine glycan array data using predefined motif lists

SignalFinder-Microarray: an image analysis tool

MCAW-DB utilizes Multiple Carbohydrate Alignment with Weights (MCAW) algorithm to align glycan structures

CCARL enables identification of terminal residues

Structural information

GlyMDB: links glycan microarray binding data from the CFG database to protein structures (PDB)

Gly-Spec (Grafting) uses structural data to predict glycan microarray binding

Grace Microarray products for Glycan analysis:

ONCYTE® Film slides

PATH® Protein Microarray Slides

Epoxy Microarray Slides

Microarray Reagents

ProPlate® 

Referrences:

Mehta AY, Heimburg-Molinaro J, Cummings RD. Tools for generating and analyzing glycan microarray data. Beilstein J Org Chem. 2020;16:2260-2271. Published 2020 Sep 10. doi:10.3762/bjoc.16.187