Deruxtecan (Dxd) ADC ELISA Kit

Sample

serum, plasma

Species Reactivity

N/A

Detection Method

sELISA

Intended Use

Creative Diagnostics has comprehensively developed and validated the DXd ADC ELISA Kit for the quantitative measurement of DXd-ADC in serum and plasma.

Contents of Kit

The kit provides the following reagents and solutions for the quantitative measurement of DXd-ADC in biological matrices.
1. Capture Plate, 1 plate.
2. Standard Stock, 1 vial (20 μL)
3. 100× Detection Antibody [HRP], 1 vial (150 μL)
4. Sample Dilution Buffer, 1 bottle (60 mL)
5. 20× Wash Solution, 1 bottle (60 mL)
6. Stop Solution, 1 bottle (6 mL)
7. TMB Solution, 1 bottle (12 mL)
8. Plate Sealer, 2 pieces
Capture Plate: 96 well microplates (8 wells × 12 strips); 12 strips are configured in plate; plate is sealed in a foil pouch with a desiccant.
Standard Stock contains 100 μg/mL of DXd-ADC.

Storage

The unopened kit is stable for 12 months from the date of manufacture at 2°C to 8°C, and the opened kit is stable for 30 days from the date of opening at 2°C to 8°C.

Performance Characteristics

Selectivity
Selectivity was tested by spiking plasma of ten different samples from non-human primate with DXd-ADC Quality Controls at two concentrations (HQC of 960 ng/mL and LLOQ of 20 ng/mL). Based on the ICH M10 guidance, the mean accuracy for LLOQ was required to be within 75%-125% of the low spiked concentration in at least 90% of the evaluated matrices. The mean accuracy for HQC was required to be within 80%-120% of the high spiked concentration in at least 100% of the evaluated matrices (Table).
Selectivity analysis of the Kit:

Precision

DXd-ADC Quality Controls at three concentrations (HQC of 960 ng/mL, MQC of 192 ng/mL, and LQC of 60 ng/mL) were tested for precision and accuracy in three batches of the kits, with each sample tested three times.
Intra-assay and inter-assay precision and accuracy of the kit:

General Description

DXd-ADC technology is composed of an enzymatically cleavable tetrapeptide–based linker, a novel exatecan derivative (DXd) payload and an antibody drug. For example, Trastuzumab deruxtecan (DS-8201a, T-DXd) is an DXd antibody-drug conjugate (DXd-ADC), composed of an enzymatically cleavable maleimide glycynglycyn-phenylalanyn-glycyn (GGFG) peptide linker, DXd, and an anti-HER2 antibody. DXd is a more potent DNA topoisomerase I (TOP1) inhibitor which has been proved to be cytotoxic to human cancer cell lines, such as KPL-4, NCI-N87, SK-BR-3, and MDA-MB-468. The DXd-ADC technology has a linker stable in plasma, a payload with a short systemic half-life, and an ADC in which the average drug-to-antibody ratio (DAR) can be optimized up to 8 for each target.
Creative Diagnostics has comprehensively developed and validated the DXd ADC Pharmacokinetic ELISA Kit for the quantitative measurement of DXd-ADC in cynomolgus monkey serum and plasma.

Standard Curve

A set of DXd-ADC calibration standards were freshly prepared and analyzed. Standard curves were constructed using a four-parameter logistic curve. The typical dynamic range of the kit is 20-1,280 ng/mL (0.5-32 ng/mL diluted), and its detection limit is 20 ng/mL (Table & Figure).
Sample data for standard curve:


Figure: DXd ADC ELISA Kit standard curve
A set of DXd-ADC calibration standards from 1,280 ng/mL to 20 ng/mL was then diluted with Sample Dilution Buffer with a volume ratio of 1:40.