| Product Description |
DXd-ADC technology is composed of an enzymatically cleavable
tetrapeptide–based linker, a novel exatecan derivative (DXd) payload and an antibody drug [1–3].
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 [2]. 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 [1–6].
GenScript has comprehensively developed and validated the DXd ADC
Pharmacokinetic ELISA Kit for the quantitative measurement of DXd-ADC in
cynomolgus monkey serum and plasma. This kit based
on the ICH M10 and the FDA bioanalytical method
validation guidance for industry, ensuring its precision, accuracy,
dilutional linearity, specificity, selectivity, stability, and hook effect were
acceptable [7-10]. The ELISA kit is a validated tool for DXd-ADC
quantification in biological matrices for drug research and development. |
| LLOQ |
20 ng/mL
|
| ULOQ |
1,280 ng/mL
|
| Precision |
Intra-assay: CV≤10%
Inter-assay: CV≤15%
|
| Minimum required dilution (MRD) |
1:40, validated non-human primate
plasma
|
| Kit Contents |
| Component |
Quantity/Size |
Part No. |
| Capture Plate |
1 plate |
N1-80 |
| Standard Stock |
1 vial (20 μL) |
N1-10 |
| 100× Detection Antibody [HRP] |
1 vial (150 μL) |
N1-20 |
| Sample Dilution Buffer |
1 bottle (60 mL) |
N1-60 |
| 20× Wash Solution |
1 bottle (60 mL) |
N1-70 |
| Stop Solution |
1 bottle (6 mL) |
A1-50 |
| TMB Solution |
1 bottle (12 mL) |
A1-40 |
| Plate Sealer |
2 pieces |
N/A |
|
| Storage |
The
unopened kit is stable for at least 12 months from the date of manufacture at
2°C to 8°C, and the opened kit is stable for up to 30 days from the date of
opening at 2°C to 8°C. |
| Assay Principle |
DXd ADC Pharmacokinetic ELISA Kit is a
sandwich ELISA assay that utilizes an anti-DXd monoclonal antibody as the capture antibody
and an anti-human IgG monoclonal antibody as the detection antibody. When
standards or samples are added to the capture plate, the anti-DXd monoclonal
antibody coating on the plate can capture the DXd-ADC present in the sample. Then
the Horseradish Peroxidase (HRP) conjugated Anti-human IgG monoclonal antibody is
added to interact with the DXd-ADC bound on the plate.
After the washing steps, 3,3',5,5'-Tetramethylbenzidine solution (TMB Solution)
is added, resulting in the formation of blue color. The reaction is stopped by
adding Stop Solution. Adding the Stop Solution changes the color from blue to
yellow. The intensity of the color can be read at 450 nm and 630 nm by a
microplate reader. The quantity of DXd-ADC in the sample is accurately
determined against a DXd-ADC standard curve. |
| Reference |
1. Nakada T, Masuda T, Naito H, Yoshida M, Ashida S, Morita K, et al. Novelantibody drug conjugates containing exatecan derivative-based cytotoxicpayloads. Bioorganic Med Chem Lett 2016; 26:1542–5.
2. Ogitani Y, Aida T, Hagihara K,Yamaguchi J, Ishii C, Harada N, et al. DS-8201a, a novel HER2-targeting ADCwith a Novel DNA topoisomerase I inhibitor, demonstrates a promising antitumorefficacy with differentiation from T-DM1. Clin Cancer Res 2016; 22:5097–108.
3. Nakada T, Sugihara K, Jikoh T, Abe Y, Agatsuma T. The latest research anddevelopment into the antibody–drug conjugate, [fam-] trastuzumab deruxtecan(DS-8201a), for HER2 cancer therapy. Chem Pharm Bull 2019; 67:173–85.
4. Hashimoto Y, Koyama K, Kamai Y, Hirotani K, Ogitani Y, Zembutsu A, et al.A novel HER3-targeting antibody–drug conjugate, U3–1402, exhibits potenttherapeutic efficacy through the delivery of cytotoxic payload by efficientinternalization. Clin Cancer Res 2019; 25:7151–61.
5. Iida K, Abdelhamid Ahmed AH, Nagatsuma AK, ShibutaniT, Yasuda S, Kitamura M, et al. Identification and therapeutic targeting ofGPR20, selectively expressed in gastrointestinal stromal tumors, with DS-6157a,a first-in-class antibody-drug conjugate. Cancer Discov 2021; 11:1508–23.
6. Okajima D, Yasuda S, Maejima T, Karibe T, Sakurai K, Aida T, et al.Datopotamab deruxtecan, a novel TROP2-directed antibody–drug conjugate,demonstrates potent antitumor activity by efficient drug delivery to tumorcells. Mol Cancer Ther 2021; 20:2329–40.
7. International Council for Harmonisation of Technical Requirements forPharmaceuticals for Human Use. ICH Harmonised Guideline, Bioanalytical MethodValidation M10 (2019).
8. US FDA. Bioanalytical Method Validation Guidance for Industry (2018).
9. European Medicines Agency. Guideline on Bioanalytical Method Validation(2011).
10. DeSilva B, Smith W, Weiner R et al. Recommendationsfor the bioanalytical method validation of ligand-binding assays to support.
|
Figure 1: DXd ADC Pharmacokinetic 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.
For research use only.
Not intended for human and animal therapeutic or diagnostic use.
