| Product Description |
Monomethyl auristatin E (MMAE) is a synthetic antineoplastic
agent. It inhibits cell division by blocking the polymerisation of tubulin.It is linked to a monoclonal antibody as a payload in MMAE
conjugated antibody drug (MMAE-ADC) development
[1-3]. There are several FDA approved MMAE-ADC drugs, such as Disitamab
vedotin, Tivdak, Padcev, Polivy and Adcetris. MMAE-ADC can activate
the antimitotic mechanism by cathepsin cleavage after it has entered a tumor
cell, thus [4].
GenScript has comprehensively developed and
validated the MMAE ADC
Pharmacokinetic ELISA Kit for the quantitative measurement of MMAE-ADC in
cynomolgus monkey serum and plasma. This kit is 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
[5-8]. The ELISA kit is a validated tool for MMAE-ADC quantification
in biological matrices for drug research and development. |
| LLOQ |
15.6 ng/mL
|
| ULOQ |
1,000 ng/mL
|
| Precision |
Intra-assay: CV≤10%
Inter-assay: CV≤15%
|
| Minimum required dilution (MRD) |
1: 20, validated non-human
primate plasma
|
| Kit Contents |
| Component |
Quantity/Size |
Part No. |
| Capture Plate |
1 plate |
S1-80 |
| Standard Stock |
1 vial (20 μL) |
S1-10 |
| 100× Detection Antibody [HRP] |
1 vial (150 μL) |
S1-20 |
| Sample Dilution Buffer |
1 bottle (60 mL) |
S1-60 |
| 20× Wash Solution |
1 bottle (60 mL) |
S1-70 |
| TMB Solution |
1 bottle (12 mL) |
A1-40 |
| Stop Solution |
1 bottle (6 mL) |
A1-50 |
| 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 1 month from the date of
opening at 2°C to 8°C. |
| Assay Principle |
MMAE ADC Pharmacokinetic ELISA Kit is a
sandwich ELISA assay that utilizes an anti-MMAE 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-MMAE monoclonal antibody coating on the plate can
capture the MMAE-ADC present in the sample. Then the Horseradish Peroxidase
(HRP) conjugated Anti-human IgG monoclonal antibody is added to interact with the MMAE-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 MMAE-ADC in the sample is accurately determined against a MMAE-ADC standard curve. |
| Reference |
1. Francisco JA, Cerveny CG, Meyer DL, et al. cAC10-vcMMAE, ananti-CD30-monomethyl auristatin E conjugate with potent and selective antitumoractivity. Blood. 2003 Aug 15;102(4):1458-65. Epub 2003 Apr 24.
2. Junutula JR, Raab H, Clark S, et al. Site-specificconjugation of a cytotoxic drug to an antibody improves the therapeutic index.Nat Biotechnol. 2008 Aug;26(8):925-32.
3. Doronina SO, Toki BE, Torgov MY, et al. Development of potent monoclonalantibody auristatin conjugates for cancer therapy. Nat Biotechnol. 2003Jul;21(7):778-84.
4. Joseph A Francisco 1, Charles G Cerveny, Damon L Meyer, et al. cAC10-vcMMAE,an anti-CD30-monomethyl auristatin E conjugate with potent and selectiveantitumor activity. Blood 2003 Aug 15;102(4):1458-65.
5. International Council for Harmonisation of Technical Requirements forPharmaceuticals for Human Use. ICH Harmonised Guideline, Bioanalytical MethodValidation M10 (2019).
6. US FDA. Bioanalytical Method Validation Guidance for Industry (2018).
7. European Medicines Agency. Guideline on Bioanalytical Method Validation(2011).
8. DeSilva B, Smith W, Weiner R et al. Recommendationsfor the bioanalytical method validation of ligand-binding assays to support. |
Figure 1: MMAE ADC Pharmacokinetic ELISA Kit standard curve
A set of MMAE-ADC calibration standards from 15.6 ng/mL to 1000 ng/mL was then diluted with Sample Dilution Buffer with a volume ratio of 1:20.
For research use only.
Not intended for human and animal therapeutic or diagnostic use.
