Therapeutic Drug Monitoring ELISA Assay

Vedolizumab Antibodies ELISA Assay

$1,485.00

The Vedolizumab Antibodies ELISA Assay is an enzyme immunoassay for the semi-quantitative determination of  free antibodies to Vedolizumab (Entyvio) in serum and plasma. The Eagle Biosciences Vedolizumab (Entyvio®) Antibodies ELISA Assay Kit is for research use only and is not intended for diagnostic or therapeutic procedures.

SKU: IG-BB116 Categories: , , ,

Vedolizumab Antibodies ELISA Assay

Vedolizumab Antibodies ELISA Assay for Research Use Only

Size: 1×96 wells
Sensitivity: Cut-Off
Incubation Time: 2.5 hours
Sample Type: Serum, Plasma
Sample Size: 10 µL
Alternative Names: Entyvio Assay

Assay Principle

This anti-drug antibody(ies) (ADA) kit is a bridging type ELISA for the determination of free antibodies against the drug Vedolizumab in serum and plasma samples. During the first incubation period, ADA in serum or plasma samples are captured by the drug coated to the microtiter wells. After washing away the unbound components from samples, a peroxidase-labelled drug conjugate is added and then incubated. ADA, if present in the sample, will make a bridge, with its identical Fab arms, between the drug coated on the well and the other drug molecule labelled with peroxidase. After a second washing step, the bound enzymatic activity is detected by addition of tetramethylbenzidine (TMB) chromogen-substrate. Finally, the reaction is terminated by adding a stop solution. The positive reaction is expected to be related to the presence of ADA in the sample.

STORAGE AND STABILITY OF THE KIT
The Vedolizumab Antibodies ELISA Assay kit is shipped at ambient temperature and should be stored at 2-8°C. Keep away from heat or direct sun light. The storage and stability of specimen and prepared reagents is stated in the corresponding chapters. The microtiter strips are stable up to the expiry date of the kit in the broken, but tightly closed bag when stored at 2–8°C.

Related Products

Vedolizumab mAb-based ELISA
Total Antibodies to Infliximab (Remicade®) ELISA Kit
Nivolumab (Opdivo) Antibodies ELISA Assay

Additional Information

Assay Background

The drug Vedolizumab (trade name Entyvio®) is a humanised immunoglobulin G1 monoclonal antibody that binds exclusively to the lymphocyte integrin α4β7, is indicated for the treatment of moderately to severely active ulcerative colitis [UC] or Crohn’s disease [CD]. During treatment the presence of persistent anti-Vedolizumab antibody was observed to substantially reduce serum concentrations of Vedolizumab, either to undetectable or negligible levels.

The Eagle Biosciences Antibody to Vedolizumab ELISA Assay kit has been designed for the measurement of free antibodies against this drug. It does not detect such antibodies which already are bound to the drug.

Assay Procedure

  1. Pipette 100 µL of Assay Buffer into each of the wells to be used.
  2. Pipette 50 µL of each Ready-to-Use Negative Control, Ready-to-Use Positive Control, and 1:10 Diluted Samples (as described in section 10.2) into the respective wells of the microtiter plate. Bubble formation during the pipetting of standards and samples must be avoided.
  3. Cover the plate with adhesive seal. Shake plate carefully by tapping several times. Incubate the plate on bench top for 60 min at room temperature (RT, 20-25°C).
  4. Remove adhesive seal. Aspirate or decant the incubation solution. Wash the plate 3 X 300 μL of Diluted Wash Buffer per well. Remove excess solution by tapping the inverted plate on a paper towel.
  5. Pipette 100 μL of Enzyme Conjugate (HRP-drug) into each well.
  6. Cover the plate with adhesive seal. Shake plate carefully by tapping several times. Incubate the plate on bench top for 60 min at RT.
  7. Remove adhesive seal. Aspirate or decant the incubation solution. Wash the plate 3 X 300 μL of Diluted Wash Buffer per well. Remove excess solution by tapping the inverted plate on a paper towel.
  8. Pipette 100 µL of Ready-to-Use TMB Substrate Solution into each well.
  9. Incubate 15 min at RT. Avoid exposure to direct sunlight.
  10. Stop the substrate reaction by adding 100 µL of Stop Solution into each well. Briefly mix contents by gently shaking the plate. Color changes from blue to yellow.
  11. Measure optical density (OD) with a photometer at 450 nm (Reference at OD620 nm is optional) within 15 min after pipetting the Stop Solution.

Documents

Product Manual

 

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.

Product Citations