DNA Photoaffinity Labeling-based DEGL Screening
Overview of DNA Photoaffinity Labeling-based DEGL Screening
DNA photoaffinity labeling-based DNA-encoded glycan library (DEGL) screening combines DEGL with photoaffinity labeling techniques to identify glycan-binding interactions with high specificity and stability. This method utilizes photoactivatable crosslinkers attached to DNA-encoded glycans, which, upon exposure to UV light, form covalent bonds with nearby target molecules.
Lock in Glycan Interactions with DNA Photoaffinity Labeling-based DEGL Screening
At CD BioGlyco, our DNA photoaffinity labeling-based DEGL screening is an advanced technology designed to uncover complex glycan interactions with high specificity and stability. By integrating DNA-encoded libraries with photoaffinity labeling, this service provides researchers with a powerful tool to identify glycan-binding partners crucial for drug discovery and understanding glycan-related biological processes. Here are our standard steps for DNA photoaffinity labeling-based DEGL screening.
Workflow
Library construction
A diverse library of glycans is synthesized and each glycan is conjugated with a unique DNA barcode. Each glycan is also linked to a photoactivatable crosslinker. These crosslinkers are designed to form covalent bonds with nearby molecules upon exposure to UV light, ensuring that the glycan-target interactions are captured and stabilized.
Target incubation
The DEGL is incubated with the target of interest (e.g., proteins, receptors, or other biomolecules). During this incubation, specific glycans in the library interact with the target, forming glycan-target complexes. Non-specific interactions are removed through a series of washing steps, leaving only the glycans that have formed stable interactions with the target.
Photoactivation
The glycan-target complexes are exposed to UV light, which activates the photoaffinity labels. This causes the formation of covalent bonds between the glycans and their binding partners, permanently linking the glycans to the targets.
Isolation and PCR amplification
These isolated DNA sequences are then amplified using PCR. The amplification process increases the quantity of DNA corresponding to the bound glycans, enabling easier identification in subsequent steps.
High-throughput sequencing
The amplified DNA is subjected to high-throughput sequencing, and the sequencing data is analyzed to determine which glycans have formed stable interactions with the target.
Validation and functional studies
Identified glycan interactions are validated through additional biochemical assays or functional studies to confirm their relevance and potential as therapeutic targets. The validated interactions can be further explored for drug development, biomarker discovery, or mechanistic studies in glycan-related pathways.
Applications
- Our service is particularly useful for identifying glycan-binding targets that play crucial roles in disease pathways, making it a valuable tool in the early stages of drug discovery.
- The ability to identify and validate glycan interactions with high specificity allows for the discovery of novel biomarkers.
- Glycans are key components of many pathogens' surfaces. This service can help identify glycans that interact with immune receptors, paving the way for the development of glycan-based vaccines or adjuvants that enhance immune responses.
- Researchers can use this service to explore the functional roles of glycans in various biological processes, including cell signaling, immune modulation, and pathogen-host interactions.
Advantages
- The integration of DNA encoding and high-throughput sequencing allows for the simultaneous screening of vast libraries of glycans, accelerating the discovery of significant interactions and reducing the time required for research and development.
- By providing a streamlined and efficient method for identifying glycan interactions, this service reduces the resources needed for traditional screening methods, making it a cost-effective solution for both academic and industrial research.
- Our service offers detailed insights into glycan-target interactions, including high-resolution data that can be used for further functional studies, validation, and application in therapeutic development.
Publication
Technology: Photo-affinity labeling, Click chemistry, SDS-PAGE, Fluorescence imaging, Pull-down assays, LC-MS/MS, ATP production assays
Journal: PLoS Neglected Tropical Diseases
IF: 3.4
Published: 2017
Results: The study successfully identified the mitochondrial FoF1-ATP synthase (complex V) as the target of the bis-tetrahydropyran 1,4-triazole (B-THP-T) compounds in Trypanosoma brucei. Using a bi-functional photo-affinity probe, the research demonstrated that these compounds inhibit ATP production by specifically targeting the α- and β-subunits of the FoF1-ATP synthase, thus blocking oxidative phosphorylation. This discovery paves the way for the further development of trypanocidal drugs targeting this essential enzyme in the parasite.
Fig.1 Labelling and detection of B-THP-T target(s). (Tulloch, et al., 2017)
Frequently Asked Questions
- How does photoaffinity labeling enhance the screening process?
- Photoaffinity labeling creates stable covalent bonds between glycans and their targets upon UV activation, ensuring that the interactions are permanently captured. This stability allows for more accurate detection and analysis of glycan-target interactions.
- Can I customize the glycan library for my specific research needs?
- Yes, CD BioGlyco offers the option to customize the glycan library to meet specific research needs, enabling researchers to focus on particular glycans or explore a broader range of interactions.
- What kind of outputs can I expect from this service?
- Researchers receive a comprehensive report detailing the identified glycan-target interactions, including high-throughput sequencing data and analysis that highlights key findings relevant to the research objectives.
At CD BioGlyco, our DNA photoaffinity labeling-based DEGL screening service enables researchers to uncover critical glycan interactions that are pivotal in various biological processes and disease mechanisms. Please feel free to contact us to discover how we can support your glycan-related research and more information about our service details.
Reference
- Tulloch, L.B.; et al. Photo-affinity labelling and biochemical analyses identify the target of trypanocidal simplified natural product analogues. PLoS Neglected Tropical Diseases. 2017, 11(9): e0005886.
For research use only. Not intended for any clinical use.
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