ADC assessment

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ADC Assessment
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Introduction

• As the new generation of immunotherapy, ADCs are
  meticulously constituted bio-macromolecules with
  high potential in the treatment of cancer and
  various other diseases. The information gathered
  from in vitro analysis provides a guideline for
  ADC optimization and the downstream in vivo
  assessments.

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ADC Assessment
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Content
ADC in vitro Analysis
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Biochemical analysis

• analyze of the structure, stability, and drug
  conjugation pattern of the ADC.

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Biochemical analysis

• Structural Analysis evaluate antibody/antibody
  fragment structure to ensure its correct folding
  after conjugation.
• ADC Stability Analysis ADC biochemical analysis
  that assess the physical stability (the tendency
  for auto-fragmentation and form aggregation,
  solubility, and thermal stability), chemical
  stability (minimum payload drug release at
  non-desired pH buffer systems, correct reactivity
  of the antibody during and after the conjugation
  process), in vitro serum stability (minimum
  payload drug release in serum)
• Drug Antibody Ratio (DAR) Determination crucial
  steps in ADC biochemical analysis that assess the
  drug to antibody molar ratio and the distribution
  patter of different DAR species using advanced
  chromatography and mass spectroscopy analysis.
• Conjugation Sites Analysis provide an accurate
  map of conjugated drug-linker locations via mass
  spectroscopy along with information regarding the
  physical attachment of the unreacted drug-linker
  on the antibody.

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In vitro efficacy evaluation

• using conventional antigen-bearing cultured cell
  lines to evaluate the behavior and cytotoxicity
  of an ADC

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In vitro efficacy evaluation

• ADC Affinity Evaluation determine and compare
  the antigen and Fc receptor binding affinity of
  ADC to non-conjugated antibody using approaches
  such as ELISA, flow cytometry, or surface plasmon
  resonance (SPR).
• Internalization Assays the extent of ADC
  incorporation into the cells (internalization)
  upon binding is characterized in this stage of
  ADC in vitro efficacy analysis using ELISA or
  flow cytometry
• Cytotoxicity Assays toxicity is measured by the
  cell viability upon the application of the ADC in
  a dosage-dependent manner using
  antigen-expressing cell lines.
• ADCC and CDC Assays this set of ADC in vitro
  efficacy analysis measures the antibody dependent
  cellular cytotoxicity (ADCC) and complement
  dependent cytotoxicity (CDC) of the ADC to
  provide more detailed information regarding the
  pharmacological effects of the ADC.

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3D in vitro

• using highly innovated antigen-expressing 3D
  cultured cells to evaluate the behavior and
  cytotoxicity of an ADC for the mimicry of solid
  tumor treatment. 3D in vitro platform evaluates
  several ADC performance parameters.

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3D in vitro

• ADC Targeting Profile evaluate the targeting
  pattern, efficiency, and reflect target
  distribution in a 3D cultured cell clusters in
  mimicking of the internal environment of a solid
  tumor.
• ADC Tumor Penetration break the barrier of
  single layers in the 2D cultured cells and
  provide a three-dimensional distribution profile
  of an ADC in solid tumors.
• ADC Solid Tumor Efficacy demonstrate the
  efficacy of an ADC in a whole new regime and
  generate results more accurately resemble the
  scenarios of solid tumors.