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Product-Specific Analytical Development

Product-Specific Assays represent one of the most critical components of the preclinical testing framework. These modules are not designed to assess safety, therapeutic efficacy, or PK/PD parameters; instead, they focus exclusively on direct, analytical, and mechanistic evaluation of the drug itself.

These assays investigate the intrinsic properties of the molecule, including:

  • how biologically active the drug is?
  •  whether it truly binds to its intended target?
  •  how stable it remains under various physicochemical conditions?
  •  and how much of its therapeutic activity can be demonstrated at the cellular level?

Thus, while Safety Assays reveal how the body reacts to the drug, and Efficacy Assays demonstrate what the drug does to the disease, Product-Specific Assays define the drug’s identity, quality, strength, stability, and mechanistic behavior in a direct and scientifically quantifiable manner.

The outputs of this package consist of six essential analytical modules

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    1

    Custom Drug Quantification Method Development

     This module is required when the molecule is novel, peptide-based, nanoparticle-based, or non-antibody in nature, meaning no commercial assay or established global method exists for its measurement. A validated analytical method is fundamental for accurate PK/PD studies.

    Tests and activities (with reasons):

    1. Custom ELISA development (to generate antigen/antibody pairs when no commercial kit exists)
    2. HPLC or LC–MS/MS method development (for sensitive and specific quantification of novel molecules)
    3. Standard curve & calibration curve construction (to enable accurate quantitative measurement)
    4. Full method validation: linearity, sensitivity, accuracy, precision, LLOQ/HLOQ, recovery (to meet regulatory validation criteria)
    5. Matrix effect & biological stability assessment (to ensure method reliability in serum/plasma)
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    2

    Target Engagement & Pathway Modulation Assays

    These assays establish mechanistic proof that the drug interacts with the target pathway an essential requirement for validating therapeutic relevance.

    Tests (with reasons):

    1. Western Blot (p-AKT, p-ERK, STATs) (to confirm pathway activation or inhibition)
    2. ELISA / Multiplex biomarkers (to quantify downstream signaling molecules)
    3. Flow Cytometry for receptor occupancy (to measure % of receptors bound by the drug)
    4. Receptor-binding / occupancy assays (to demonstrate direct interaction with the biological target)
    5. Signal-pathway activity reporting (to map molecular alterations following drug exposure)
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    3

    Physicochemical Stability & Degradation Profiling

    Critical for peptides, proteins, nanoparticles, and sensitive small molecules. Required for quality control and regulatory submission.

    Tests (with reasons):

    1. Stress stability testing at 4°C / 25°C / 37°C (to assess temperature-dependent degradation)
    2. DLS, SDS-PAGE, SEC-HPLC (to detect aggregation and structural changes)
    3. Nanoparticle size distribution analysis (to evaluate physical stability)
    4. Degradation pathway analysis (to identify breakdown products and mechanisms)
    5. Serum/Plasma stability testing (to assess biological stability)
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    4

    Functional Potency & Biological Activity Assessment

    These assays determine whether the drug is capable of producing the expected cellular or molecular response and are core to establishing the Mechanism of Action (MoA).

    Tests (with reasons):

    1. Cell-based potency assays (to measure biological activity in functional cell models)
    2. cAMP assay (to evaluate GPCR/GLP-1 signaling activity)
    3. Luciferase reporter assays (to monitor transcriptional or inflammatory pathways)
    4. Functional cellular assays (to assess direct activation/inhibition of target pathways)
    5. Enzymatic activity assays (to measure biochemical effects mediated by the drug)
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    5

    Receptor Binding Kinetics & Affinity Measurement

    Affinity parameters (Kd, EC50, IC50) are essential for scientific publications, investor discussions, and regulatory evaluation.

    Tests (with reasons):

    1. Surface Plasmon Resonance (SPR) (to measure real-time binding kinetics)
    2. Biolayer Interferometry (BLI) (label-free affinity quantification)
    3. Radioligand binding assays (high-sensitivity receptor binding assessment)
    4. Kd, EC50, IC50 calculations (to quantify binding strength and functional potency)
    5. kon / koff kinetic analysis (to determine association/dissociation speed)