ADC PK Study

Pharmacokinetics is the study and characterization of the time course of drug absorption, distribution, metabolism, and excretion (ADME). Due to the complexity of ADCs that combine an antibody, a cytotoxic drug (also known as payload), and a specialized chemical linker, there are several analytes that need to be measured to characterize ADC PK profiles, including

total antibody, which is the ADC with a drug-antibody ratio (DAR) higher than or equal to zero

drug-conjugated antibody, which is the ADC with a DAR greater than or equal to one

free drugs (payloads) or their metabolites

ELISA immunoassay is a commonly used quantitative analytical method to measure the total and drug-conjugated antibody kinetic profiles. Multiple formats as shown in Figure 1 can be applied to detect target analytes.

Figure 1. Schematic images of ELISA-based ADC PK study

Case Study

• Drug-conjugated Antibody measurement by anti-payload antibody and anti-idiotype antibody
  

  Figure 2. Standard curve of DXd conjugated Trastuzumab Sandwich ELISA by anti-idiotype antibody and DXd Antibody. Based on drug-conjugated antibody assay format 2 in Figure 1..

  The Sandwich ELISA assay was developed using Anti-Trastuzumab Antibody (15H2), mAb, Mouse (GenScript, A02033) and DXd Antibody (39F4), mAb, Mouse (GenScript, A02217) and as the capture and detection antibodies, respectively.

  In this ELISA assay, DXd Antibody (39F4), mAb, Mouse (GenScript, A02217) was labeled with HRP. GenScript can provide customized conjugation service for this product per customer's request.

  Capture antibody: Anti-Trastuzumab Antibody (15H2), mAb, Mouse, 2 μg/ml
  Detection antibody: DXd Antibody (39F4), mAb, Mouse, 0.22 μg/ml.

• Drug-conjugated Antibody measurement by anti-payload antibody and anti-Human IgG Fc antibody
  

  Figure 3. Standard curve of DXd conjugated Trastuzumab Sandwich ELISA by anti-Human IgG Fc antibody and DXd antibody in monkey serum. Based on drug-conjugated antibody assay format 3 in Figure 1..

  The DXd conjugated Trastuzumab Sandwich ELISA assay was developed using DXd Antibody (39F4), mAb, Mouse (GenScript, A02217) and Mouse Anti-Human IgG Fc Antibody (50B4A9)[HRP], mAb (GenScript, A01854) as the capture and detection antibodies, respectively.

  The samples tested in this assay were DXd conjugated Trastuzumab. The control samples were not serum spiked. The monkey serum samples were spiked with cynomolgus monkey serum matrix.

• Total Antibody measurement by anti-idiotype antibody pairs
  

  Figure 4. Standard curve of Trastuzumab Sandwich ELISA. The Trastuzumab Sandwich ELISA assay was developed using Anti-Trastuzumab Antibody (11C4), mAb, Mouse (GenScript, A02032-40) and Anti-Trastuzumab Antibody (15H2), mAb, Mouse (GenScript, A02033-40) as the capture and detection antibodies, respectively. Based on total antibody assay format 4 in Figure 1..

  In this ELISA assay, Anti-Trastuzumab Antibody (15H2), mAb, Mouse (GenScript, A02033-40) was labeled with Biotin.

  GenScript can provide customized conjugation services for this product per the customer's request. The sensitivity of detecting Trastuzumab was observed to be up to 61 pg/ml.

Resources

ADC Analysis Tools Brochure

PK and ADA analysis tools help you better explore the process of drug metabolism and the impact of drug immune reactions

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