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DM1 Antibody Drug Conjugate (ADC) ELISA Assay Kit

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$1,850.00
SKU:
KTR-756

 Product Description

Package insert view pdf

 

 

DM1 Antibody Drug Conjugate (ADC) ELISA Assay kit: 
For Research Use Only

Size:  1 x 96 wells
Sensitivity:  0.024 µg/mL
Dynamic Range:  0.016 - 1.0 µg/mL
Incubation Time:  2.5 hours

Sample Type:  Serum, Cell Culture, Tissue
Sample Size: 100 µL
Species:  Human, Mouse, Rat, Primate

Intended Use

The Eagle Biosciences DM1 Antibody Drug Conjugate (ADC) ELISA Assay Kit (enzyme-linked immunoassay kit) is intended for use in the quantitative determination of antibody-DM1-conjugate (Trastuzumab emtansine) level in test samples. It is useful for pre-clinical and clinical pharmacology study of DM1 Antibody Drug Conjugate (ADC).

  • Samples from tissue/cell culture and serum samples from human, rat, mouse, primate, etc. can be used directly with this kit.
  • Both humanized monoclonal antibody based DM1-ADC and mouse monoclonal antibody based DM1-ADC can be measured with this ELISA Assay kit.


Assay Principle
The Eagle Biosciences DM1 Antibody Drug Conjugate (ADC) ELISA Assay Kit is designed, developed and produced for the quantitative measurement of antibody DM1 conjugate (Trastuzumab emtansine)  in serum, tissue, and cell culture samples. The assay utilizes the competitive immunoassay technique with an antibody that exclusively binds to DM1.

Assay calibrators (antibody DM1 conjugate) and test serum samples are added directly to wells of a microtiter plate that is coated with specific anti-DM1 antibody. Subsequently, a horseradish peroxidase (HRP) conjugated DM1 is added to each well. During the incubation period, the antibody DM1 conjugate competes with the HRP conjugated DM1 for the limited binding sites of anti-DM1 antibody. An immune complex of well coated “anti-DM1 antibody – HRP conjugated DM1” 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-DM1 conjugate in the test sample. A calibration curve is generated by plotting the absorbance versus the respective antibody-DM1 conjugate concentration for each calibrator on a 4-parameter or log-logit curve fitting. The concentration of antibody-DM1 conjugate in test samples is determined directly from this calibration curve.

  1. Place a sufficient number of Anti-DM1 antibody coated microwell strips/wells in a holder to run human Anti-DM1 standards, controls and unknown samples in duplicate.
  2. Add 100 µL of calibrators and test samples into the designated microwells.
  3. 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.
  4. Immediately add 25 µL of HRP Conjugated DM1 to each well. (Note: no wash step before add the HRP Conjugated DM1)
  5. 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.
  6. Wash each well 5 times by dispensing 350 µL of working wash solution into each well and then completely aspirating the contents. 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.


Typical Standard Curve:


Specificity

This DM1 ADC ELISA Assay Kit doesn’t show any cross reactivity to MMAE-ADC, MMAF-ADC, DUO-3 ADC, or DUO-6 ADC.

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

 

Recent Citations

Yao, HP;Feng, L;Zhou, JW;Zhang, RW;Wang, MH;Therapeutic evaluation of monoclonal antibody-maytansinoid conjugate as a model of RON-targeted drug delivery for pancreatic cancer treatment, Am J Cancer Res, pg. 937-56, 6:5, 2016.

 

Related News:

Antibody Drug Kits- DM1, MMAE, MMAF: Product Highlights 

New Antibody–Drug Conjugate (ADC) Study Utilizes Eagle Biosciences' DM1 ADC ELISA Assay Kit 

October 2014: Eagle Biosciences Introduces New Antibody Drug Conjugate ELISA Assays 

 

 

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Eagle Biosciences, Inc.
20A NW Blvd., Suite 112
Nashua, NH 03063
Email: info@eaglebio.com
Toll Free: 866-411-8023
International: +617-419-2019

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