MMAF Antibody Drug Conjugate (ADC) ELISA Assay Kit

$1,850.00

The Eagle Biosciences MMAF Antibody Drug Conjugate (ADC) ELISA Assay kit is intended for use in the quantitative determination of antibody-MMAF-conjugate level in test sample. It is useful for pre-clinical and clinical pharmacology study of MMAF Antibody Drug Conjugate (ADC). Samples from tissue/cell culture and serum samples from human, rat, mouse, primate, etc. can be used directly with this MMAF Antibody Drug Conjugate (ADC) ELISA Assay kit. Both humanized monoclonal antibody based MMAF-ADC and mouse monoclonal antibody based MMAF-ADC can be measured with this MMAF Antibody Drug Conjugate (ADC) ELISA Assay kit.

MMAF Antibody Drug Conjugate (ADC) ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 0.019 µg/mL
Dynamic Range: 0.016 – 2.0 µg/mL
Incubation Time: 2.5 hours
Sample Type: Serum, Cell Culture, Tissue
Sample Size: 100 µL

Product Developed and Manufactured in the USA

Additional Information

Assay Principle

The MMAF Antibody Drug Conjugate (ADC) ELISA Assay kit is designed, developed and produced for the quantitative measurement of antibody MMAF conjugate in serum. The assay utilizes the competitive immunoassay technique with an antibody that exclusively binds to MMAF.

Assay calibrators (antibody MMAF conjugate) and test serum samples are added directly to wells of a microtiter plate that is coated with specific anti-MMAF antibody. Subsequently, a horseradish peroxidase (HRP) conjugated MMAF is added to each well. During the incubation period, the antibody MMAF conjugate competes with the HRP conjugated MMAF for the limited binding sites of anti-MMAF antibody. An immune complex of well coated “anti-MMAF antibody – HRP conjugated MMAF” is formed. The unbound antibodies and buffer matrix are removed in the subsequent washing step. For the detection of this immunocomplex, the well is then incubated with a substrate solution in a timed reaction, which is terminated with an acidic reagent (i.e. ELISA stop solution). The absorbance is then measured in a spectrophotometric microplate reader. The enzymatic activity of the immunocomplex bound to the wall of each microtiter well is inversely proportional to the amount of antibody-MMAF conjugate in the test sample. A calibration curve is generated by plotting the absorbance versus the respective antibody-MMAF conjugate concentration for each calibrator on a 4-parameter or log-logit curve fitting. The concentration of antibody-MMAF conjugate in test samples is determined directly from this calibration curve.

  1. Add 100 µL of calibrators and test samples into the designated microwells.
  2. Seal the plate wells securely, cover with foil or other material to protect from light, and rotate on an ELISA plate shaker (small orbit radius) for 30 minutes at 400 to 450 rpm.
  3. Immediately add 25 µL of HRP Conjugated MMAF (cat# 30719) to each well. (Note: no wash step before add the HRP conjugated MMAF)
  4. Seal the plate wells securely, cover with foil or other material to protect from light, and rotate on an ELISA plate shaker (small orbit radius) for 2 hr. ± 10 minutes 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 ELISA HRP Substrate into each of the wells.
  8. Cover the plate with aluminum foil or other material to avoid exposure to light. Incubate plate static, at room temperature for 20 minutes.
  9. Immediately add 100 µL of ELISA Stop Solution into each of the wells. Mix gently.
  10. Read the absorbance at 450 nm.

Specificity

This MMAF ADC ELISA Assay Kit does NOT show any cross reactivity to MMAE-ADC, DM1-ADC, DUO-3-ADC, and DUO-6-ADC.

Typical Standard Curve

MMAF Antibody Drug Conjugate (ADC) ELISA Assay Kit Standard Curve

Manual

Product Manual



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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