Overview of the Importance of Hit Synthesis and Activity Verification

Hit compound synthesis and activity verification are key steps in constructing a DNA-encoded Glycan Library (DEGL). Initial hit compounds identified from the screening process are resynthesized to ensure that the observed activity is not the result of experimental artifacts or library preparation errors. After synthesis, these hit compounds undergo rigorous activity verification, which is testing under different conditions to confirm their binding affinity, specificity, and biological activity. This two-layer approach not only filters out false positives but also improves the reliability and stability of the discovered glycan ligands. Ensuring precise synthesis and accurate activity verification ultimately identify truly effective and relevant glycan structures, thereby facilitating the development of therapeutic agents and diagnostic tools. At CD BioGlyco, we provide clients with Hit and Analogue Synthesis, hit activity verification, and Hit Structure-activity Relationship (SAR) Research services based on our years of experience and rich knowledge accumulation.

Ensuring Precision, Powering Discovery: DNA-Encoded Glycan Hit Activity Verification

At CD BioGlyco, our hit activity validation service involves several precise and methodical steps to ensure accurate and reliable hit validation. Our service process is as follows:

Preparation and Encoding Services of DEGL

DEGL preparation and encoding is the systematic synthesis and labeling of glycans using unique DNA sequences as molecular barcodes. The process begins with chemical synthesis or extraction of glycans, followed by the binding of each glycan to a specific DNA sequence through a powerful chemical reaction, ensuring precise encoding. The DNA tag serves a dual purpose: it acts as a unique identifier for each glycan, allowing for high-throughput screening (HTS) and easy tracking. The DNA tag provides a method for amplification and sequencing, which is essential for downstream analysis and deconvolution of screening results. Our meticulous preparation and encoding services ensure that each glycan-DNA conjugate is accurately represented and is reliably identified and analyzed in complex bioassays.

DEGL Screening and Binding Assay Services

This step is to systematically evaluate the interaction between the target molecule and the compounds from DEGL. We incubate DEGL with the target to allow potential binders to form a complex. Unbound and weakly interacting molecules are then washed away to ensure specificity. The remaining bound molecules are amplified and sequenced to identify the ligands with the highest affinity. We provide precise, high-throughput, and quantitative binding affinity measurements to help clients discover glycan-target interactions with potential therapeutic applications.

Hit Identification and Synthesis Services

Hit identification allows the pinpointing of promising compounds with specific binding affinity or activity and their chemical synthesis. Bound glycans are extracted and DNA sequenced to identify glycan structures associated with positive hits. Hit compounds and analogs are synthesized and their SAR are studied to optimize their properties.

Hit Activity Verification Services

Activity validation ensures the effectiveness and potential efficacy of active compounds identified in HTS experiments. Our hit activity verification includes the following:

• Secondary assays: Use more refined and targeted assays to confirm the initial screening results. These assays often use different conditions or methods to ensure that the initial hit is not a false positive.
• Orthogonal assays: Apply completely different methods or technologies to verify hit activity, providing cross-validation to enhance the credibility of research results.
• Dose-response studies: Evaluate the activity of compounds at different concentrations to determine potency.
• Physicochemical characterization: Analyze the stability, solubility, and other physical properties of the compound to ensure that the observed activity is not due to degradation or aggregation of the compound.
• Selectivity and specificity testing: Target validation analysis, off-target screening and a large number of potential interaction analyses confirm the specificity of the target compound for the intended target.
• Cell and in vivo validation: Test the target compound in cell models or animal experiments to determine its biological relevance and efficacy in more complex physiological environments.

Workflow

Applications

• Hit activity verification ensures the integrity and reliability of initial screening results by confirming the bioactivity of active compounds identified from HTS or other discovery approaches. This process reduces false positives and helps identify truly active molecules.
• This provides detailed insight into the mechanism of action of active compounds. By validating these active compounds through a variety of bioassays, researchers gain insight into how these compounds interact with their target proteins or pathways.
• By evaluating the bioactivity of a range of analogs, scientists analyze key structural features that lead to activity and design more potent and selective compounds.
• Bioactivity validation informs chemical modifications needed to improve efficacy, reduce off-target effects, and enhance pharmacokinetic properties.

Advantages

• We use advanced methods such as confirmatory assays, secondary screening, and statistical analysis to ensure that truly promising compounds progress in the drug discovery process. This enables more efficient, cost-effective research and development and accelerate the identification of viable therapeutic candidates.
• We have experienced experts to guide us throughout the entire process, which provides significant advantages for the accuracy and reliability of data interpretation in drug activity verification services. This reduces the possibility of false positives and false negatives, saving time and resources in subsequent development phases.
• Our verification process strictly adheres to compliance standards to provide our clients with reliable and timely information to make informed decisions.
• Our commitment to continuous improvement and innovation keeps us at the forefront of the industry, providing unparalleled service quality and customer satisfaction.

Publication Data

Frequently Asked Questions

• How do you verify hit activity?
  • Hit activity is typically verified through a series of secondary assays and orthogonal validation techniques. These include:
  • Repeating the initial assay to check for reproducibility.
  • Performing dose-response studies to determine the potency and efficacy of the hits.
  • Using alternative complementary assays to confirm the activity.
  • Evaluating the selectivity of the hits against a range of targets to ensure specificity.
  • Cross-validation with similar yet different experimental setups or detection methods.
• Can SAR analysis be part of hit activity verification?
  • Yes, SAR analysis often plays a role in hit verification by exploring how structural modifications to the hit compound impact its biological activity. This helps in refining and optimizing lead compounds with improved properties.

At CD BioGlyco, our hit activity verification service is designed to ensure the reliability and validity of potential hits identified from DEGLs. Our comprehensive service involves a complex validation process, including rigorous binding assays, biochemical activity assessments, and structural validation techniques. Please feel free to contact us to learn about the process, strategy, and technology of our hit activity verification service.

• Hit and Analogue Synthesis
• Structure-Activity Relationship (SAR) Optimization of Hits