Intact MMAE ADC ELISA

$1,965.00

The Intact MMAE ADC ELISA Assay Kit is a highly sensitive “sandwich” test kit is intended for use in the quantitative determination of antibody MMAE conjugate level in serum or plasma of human, mouse, rat, primate, etc. It is useful for pre-clinical and clinical pharmacology study of MMAE Antibody Drug Conjugate (ADC). The Eagle Biosciences Intact MMAE ADC ELISA Assay Kit is for research use only and not used for diagnostic purposes.

SKU: MME91-K01 Categories: , ,

Intact MMAE ADC ELISA

Intact MMAE ADC ELISA Developed and Manufactured in the USA

Size: 1×96 wells
Sensitivity: 1.12 ng/mL
Dynamic Range: 54.8 – 390.0 ng/mL
Incubation Time: 2 hours
Sample Type: Serum, Plasma
Sample Size: 10 µL
Alternative Names: Intact Monomethyl auristatin E ADC ELISA, Intact MMAE Antibody Drug Conjugate
For Research Use Only

Controls Included


Assay Principle

The Intact MMAE ADC ELISA is designed, developed and produced for the quantitative measurement of antibody MMAE conjugate in serum or plasma. The assay utilizes the sandwich immunoassay technique with an antibody that binds to MMAE. Briefly, a mouse monoclonal antibody specific to MMAE is coated onto a microtiter plate.

In the assay system, the assay calibrators, controls and test specimen are added to this microtiter plate. During the first incubation period, the anti-MMAE monoclonal antibody captures the MMAE-Antibody Conjugate of calibrators, controls and test samples. Unbound proteins are washed away with a wash step. A HRP (horseradish peroxidase) conjugate anti-human IgG tracer antibody is added to each well of the microtiter plate. After the second incubation, a “sandwich” immunocomplex of “Anti-MMAE antibody – MMAE Antibody Conjugate – HRP-conjugated anti-human IgG antibody” is formed and attached to the wall of the plate. The unbound HRP-conjugated antibody is removed in a subsequent washing step. For the detection of this immunocomplex, each well is then incubated with a substrate solution in a timed reaction and then measured in a spectrophotometric microplate reader. The enzymatic activity of the immunocomplex bound to MMAE Antibody Conjugate on the wall of the microtiter well is directly proportional to the amount of MMAE Antibody Conjugate level in the sample.


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Additional Information

Assay Procedure


  1. Bring all reagents to room temperature (18-25°C) before use. Mix gently without causing foam.
  2. Add 25 µL of calibrators and diluted 1:100 test samples into the designated microwells. Tap the plate gently.
  3. Immediately add 100 µL of Assay Buffer
  4. Seal the plate wells securely, cover with foil or similar material to protect from light, and rotate on an ELISA plate shaker (small orbit radius) for 1 hour at 400 to 450 rpm.
  5. Wash each well 5 times by dispensing 350 µL of working wash solution into each well and then completely aspirating the contents.
  6. Alternatively, an automated microplate washer can be used.
  7. Add 100 µL of MMAE Tracer Antibody to each well. Tap the plate gently.
  8. Seal the plate wells securely, cover with foil or similar material to protect from light, and rotate on an ELISA plate shaker (small orbit radius) for 30 minutes at 400 to 450 rpm.
  9. Wash each well 5 times by dispensing 350 µL of working wash solution into each well and then completely aspirating the contents.
  10. Alternatively, an automated microplate washer can be used.
  11. Add 100 µL of ELISA HRP Substrate into each of the wells.
  12. Cover the plate with aluminum foil or similar material to avoid exposure to light. Incubate plate static, at room temperature for 20 minutes.
  13. Immediately add 100 µL of ELISA Stop Solution into each of the wells. Mix gently.
  14. Read the absorbance at 450 nm with reference filter at 620 nm.

Typical Standard Curve


Intact MMAE ADC ELISA

Specificity


This Eagle Biosciences Intact MMAE ADC ELISA Assay Kit doesn’t show any cross reactivity to DM1-ADC. This assay does cross react to MMAF-ADC.   The Eagle Biosciences Intact MMAE ADC ELISA Assay Kit is a highly sensitive “sandwich” test kit is intended for use in the quantitative determination of antibody MMAE conjugate level in serum or plasma of human, mouse, rat, primate, etc.

Documents

Product Documents


 

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

Publications

References


Sandhya Girish, et al. Clinical pharmacology of trastuzumab emtansine (T-DM1): an antibody–drug conjugate in development for the treatment of HER2-positive cancer. Cancer Chemother Pharmacol (2012) 69:1229–1240

Product Citations