Resin Tip-based DEGL Affinity Screening
Overview of Resin Tip-based DEGL Affinity Screening
Resin tip-based DNA-encoded glycan library (DEGL) affinity screening is a specialized high-throughput screening method that utilizes resin-packed pipette tips to isolate and identify specific glycan-protein interactions from a DEGL efficiently. This method is particularly useful in drug discovery and glycomics research, where it is crucial to identify potential glycan ligands that bind to target proteins of interest.
Unlocking Glycan Insights with Precision and Speed-Resin Tip-based DEGL Affinity Screening
Resin tip-based DEGL affinity screening is an advanced technique that streamlines the process of identifying glycan-protein interactions by integrating several key steps into a single, automated workflow. Using resin-packed pipette tips in this method allows for high specificity and efficiency in binding, washing, and eluting the glycan-protein complexes.
Workflow
Library preparation and encoding
DEGL is prepared using a split-and-pool synthesis strategy, where each glycan is tagged with a unique DNA barcode. This barcode enables precise identification of glycans throughout the screening process.
Target protein immobilization
The target protein is immobilized onto the resin within the pipette tips, allowing for specific glycan-protein interactions to occur. The resin provides a high surface area for protein binding, ensuring that even low-abundance glycan interactions can be detected.
Washing
After incubation, a series of stringent washing steps is performed. These washes are designed to remove unbound or non-specifically bound glycans, leaving only those glycans that have a true affinity for the target protein.
Affinity Screening Process
- Binding: The DEGL is introduced into the resin-packed tips, where glycans with an affinity for the immobilized target protein bind to it.
- Washing: Non-specifically bound glycans are removed through a series of wash steps, which are automated to ensure consistency and reproducibility.
- Elution: The specifically bound glycans are then eluted from the resin using a controlled change in buffer conditions, such as pH or ionic strength, which disrupts the glycan-protein interactions.
DNA sequencing and data analysis
The DNA barcodes from the eluted glycans are amplified via PCR and sequenced using high-throughput methods such as next-generation sequencing (NGS). The sequencing data is analyzed to identify the specific glycans that are bound to the target protein, providing insights into potential therapeutic targets.
Applications
- Resin tip-based DEGL affinity screening enables the discovery of glycan ligands that interact with specific protein targets, which can be developed into novel therapeutic agents for a wide range of diseases, including cancer, infectious diseases, and autoimmune disorders.
- The technology can be used to study the interactions between glycans and proteins, providing insights into the role of glycosylation in various biological processes and diseases.
- By screening DEGLs against proteins implicated in specific diseases, this method can help identify glycan structures that serve as biomarkers for early disease research.
- Resin tip-based DEGL affinity screening can help in identifying the specific binding sites of glycans on target proteins, providing valuable information for the design of glycan-based drugs and inhibitors.
Advantages
- The use of resin tips allows for the integration of the entire process into automated systems, enabling the screening of thousands of interactions in parallel.
- The automated nature of the process ensures high precision in the handling of samples, reducing variability and increasing the reproducibility of results.
- This method can easily be scaled up for large-scale screenings, making it ideal for drug discovery applications where large libraries need to be screened quickly and efficiently.
Publication
Technology: NGS, DNA-compatible reaction, DNA-encoded chemical library
Journal: SLAS DISCOVERY: Advancing the Science of Drug Discovery
IF: 2.7
Published: 2020
Results: The research results summarized in the document focus on the effectiveness of DNA-encoded chemical libraries (DELs) as a powerful technology platform for the discovery of small-molecule ligands to protein targets of biological and pharmaceutical interest. The study highlights that DELs enable the construction and screening of vast libraries with unprecedented size, leading to the identification of potent ligands, some of which have advanced to clinical trials. The research also discusses diverse approaches for generating and screening DEL molecular repertoires and presents successful case studies. The review emphasizes recent advancements in the field and outlines future challenges, particularly in improving DEL technology for therapeutic discovery.
Fig.1 (A) Standard workflow for sample preparation of phosphopeptide samples for MS analysis. (B) General description of dSPE in the IMCStips. (Mullis, et al., 2020)
Frequently Asked Questions
- How does resin tip-based DEGL affinity screening service?
- The screening process involves binding target proteins to resin within pipette tips, incubating these with the DEGL to allow specific interactions, and then washing away non-binders. The bound glycans are eluted and identified through DNA sequencing, revealing which glycans interact with the target protein.
- How is resin tip-based DEGL affinity screening different from traditional screening methods?
- Unlike traditional methods, resin tip-based DEGL affinity screening is fully automated, reducing the risk of human error and increasing throughput. It allows for the screening of vast libraries in a fraction of the time, with greater precision and reproducibility.
At CD BioGlyco, our resin tip-based DEGL affinity screening is a versatile and powerful tool with wide-ranging applications in drug discovery, glycomics, biomarker identification, structural biology, and high-throughput screening, making it an invaluable resource for advancing research in these areas. Please feel free to contact us to discover how we can support your research and more information.
Reference
- Mullis, B.T.; et al. Automating complex, multistep processes on a single robotic platform to generate reproducible phosphoproteomic data. SLAS DISCOVERY: Advancing the Science of Drug Discovery. 2020, 25(3): 277-286.
For research use only. Not intended for any clinical use.
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