Overview

Glycans interact with glycan-binding proteins (GBPs) and play important roles in recognition, adhesion, inflammation, and other life activities. Various methods have been developed to analyze the interactions between glycans and GBPs, such as enzyme-linked immunosorbent assay (ELISA), glycan microarrays, DNA-encoded Glycan Library (DEGL), etc. At CD BioGlyco, we have conducted in-depth studies on DEGL and finally constructed an effective strategy for glycan-GBP interaction analysis using polymerase chain reaction (PCR), next-generation sequencing (NGS), etc. With this in mind, we offer a customized DEGL screening service for glycan-protein interaction studies. We aim to screen for unique glycan structures that bind to the target amongst a wide range of glycans and maximize the acceleration of your biological research.

Fig.1 Glycans and GBP in leukocyte adhesion. (Krautter & Iqbal, 2021)

Exploring the Binding Properties of Glycans by DEGL Screening

DEGL is a highly efficient analytical method that uses conventional molecular biology techniques to probe the interactions between glycans and target proteins. Our service for this one is fully customized. After understanding your needs, our experienced research team tailors DEGL to your project criteria and performs high-throughput screening to identify glycans that bind to target proteins. The workflow includes the following:

Library Design

Based on the type and structure information of specific target proteins (antibodies, lectins, etc.), our team designs DEGLs and construction routes according to clients' needs.

Library Construction

Extensive research in synthetic carbohydrate chemistry led us to synthesize a diverse and large number of research-ready glycans. After custom synthesizing a series of glycans according to assay needs, we couple all these glycans to DNA to obtain the corresponding DNA-encoded glycans. Coupling involves coupling the glycans to DNA sequences using click chemistry, amine-responsive (NHS) cross-linking, and other methods or ligating via headers.

Library Screening

We conduct screens to analyze glycan-protein interactions by a variety of efficient means, including the use of DNA-tagged proteins in individual experiments to identify protein-binding ligands through high-throughput DNA sequencing, etc. The procedure is as follows:

• DNA-tagged proteins are incubated with DEGL. Proteins and glycans are bound in cell lysates.
• Unbound DNA-encoded glycans that are not bound to the target protein are washed away and high-fold enrichment of small-molecule ligands with high affinity is achieved.
• The binding profiles of the proteins are analyzed by amplification and NGS identification of barcode information. The number of DNA codes replicated represents the relative binding specificity of the corresponding glycan structure to the protein.

Data Analysis and Hit Validation

We have expertise in bioinformatics analysis and analyze the data in depth to analyze the binding specificity between glycans and proteins. After DNA sequencing and decoding the structure of the active compound, we re-synthesize it to perform affinity binding assays to examine glycan-protein interactions.

Workflow

We offer customized solutions for glycan-protein interaction analysis as shown in the figure:

Applications

• Glycan expression patterns are widely altered in retroviral infections, thrombosis, cancer, diabetes, neurodegenerative diseases, etc. Analyzing glycan-protein interactions by DEGL screening helps to understand the pathogenesis of these diseases.
• Analyzing glycan-protein interactions by DEGL screening helps to develop glycan-based therapies.
• Analysis of glycan-protein interactions by DEGL screening helps to study the function of glycans in various biological functions.

Advantages

• We complete the one-stop service of DEGL design, construction, screening, and result analysis for glycan-protein interaction validation.
• DEGL screening provides a multiplexed, highly sensitive, and effective method for glycan binding studies.
• A minimal sample is required to detect glycan-target protein interactions in the solution using our constructed DEGL.

Publication Data

Frequently Asked Questions

• What are the advantages of DEGL screening over glycan microarrays for analyzing glycan-protein interactions?
  • Glycan microarray technology, which immobilizes multiple glycans on slides to analyze their interactions with GBPs, is an indispensable tool. The technique has limitations in terms of automation and high-throughput analysis. DEGL is a high-throughput method to study the structure-function of different glycans. The method relies on quantitative PCR (qPCR), PCR, and NGS, providing a highly sensitive, multiplexed analytical, and accessible method with multiple advantages over other methods for glycan-protein interaction studies.
• How long does DEGL screening take?
  • Typically, DEGL screening varies by protein type and can take anywhere from a few weeks to a few months from library and protein preparation, to protein solid-loading and screening experiments and post-screening output sequencing. We will communicate a detailed DEGL screening schedule with you once we have planned your experiment.

CD BioGlyco provides a fully customized DEGL screening service for analyzing glycan-protein interactions and completes a one-stop shop from DEGL design, construction, and screening to validation. Welcome to contact us to explore glycan binding properties through DEGL screening. Our experienced screening team will assess your experimental needs and develop the most appropriate experimental protocol.

• Magnetic Bead-based DEGL Affinity Screening
• Resin Tip-based DEGL Affinity Screening
• Interaction-dependent PCR (IDPCR)-based DEGL Screening
• DNA Photoaffinity Labeling-based DEGL Screening
• Microfluidic Processor-based DEGL Screening
• Cell-based DEGL Screening
• DNA Programmed Affinity Labelling (DPAL)-based DEGL Screening
• DEGL Screening for Protein-Protein Interaction Inhibitor Discovery
• DEGL Screening for G Protein-coupled Receptor-Binding Compounds Discovery
• DEGL Screening for Protein Ubiquitination Study
• DEGL Screening for Bacterial Enzyme Study
• DEGL Screening for Antibody Target Identification
• DEGL Screening for Protein-templated Ligand Discovery