Anti-Ustekinumab ELISA Assay Kit


The Anti-Ustekinumab ELISA Assay Kit is used as an analytical tool for quantitative determination of Anti-Ustekinumab in serum, plasma and cell culture supernatant.

Anti-Ustekinumab ELISA Assay Kit

The Anti-Ustekinumab ELISA Assay Kit is For Research Use Only

Size: 12×8 wells
Sensitivity: <5 ng/mL
Standard Range: 5-320 ng/ml
Incubation Time: 2.5 hours
Sample Type: Serum, Plasma, Cell Culture Supernatants
Sample Size: 100 µL
<Alternative Name: Stelara

Assay Background

Ustekinumab is a human monoclonal antibody. It is directed against interleukin 12 and interleukin 23, naturally occurring proteins that regulate the immune system and immune-mediated inflammatory disorders. In two Phase III trials for moderate to severe psoriasis, the longest >76 weeks, ustekinumab was safe and effective. A third Phase III trial, ACCEPT, compared the efficacy and safety of ustekinumab with etanercept in the treatment of moderate to severe plaque psoriasis. This trial found a significantly higher clinical response with ustekinumab over the 12-week study period compared to high-dose etanercept. It also demonstrated the clinical benefit of ustekinumab among patients who failed to respond to etanercept. Ustekinumab is approved in Canada, Europe and the United States to treat moderate to severe plaque psoriasis. Anti-Drug Antibodies (ADA) may induce unwanted side effects in biopharmaceuticals. Hence, ADA has been subjected to increase in scrutiny by the regulatory authorities using immunogenicity safety studies. ADA has been observed in pre-clinical and clinical studies, resulting in significant changes in toxicology, pharmacokinetics and efficacy. These effects result from the generation of drug-induced (neutralizing) autoantibodies against Etanercept and can be responsible for allergic reaction, or even anaphylactic shock. This ELISA kit detects antibodies for Anti-Etanercept and may be used for monitoring immunogenicity.

Related Products

Ustekinumab ELISA Assay Kit
Ustekinumab mAb-based ELISA Assay
Anti-Adalimumab (Humira) ELISA Assay Kit

Additional Information

Assay Principle

The method employs the quantitative sandwich enzyme immunoassay technique. Ustekinumab is pre-coated onto microwells. Samples and standards are pipetted into microwells and antibodies to Ustekinumab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated Ustekinumab is pipetted and incubated. After washing microwells in order to remove any nonspecific binding, the ready to use substrate solution (TMB) is added to microwells and color develops proportionally to the amount of Anti-Ustekinumab in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.

Assay Procedure

  1. It is strongly recommended that all Controls and Samples be run in duplicates or triplicates. A standard curve is required for each assay. All steps must be performed at 37°C
  2. Pipette out 25 μl of Assay Diluent in each well.
  3. Pipette 25 μl of Ustekinumab: HRP Conjugate into each well.
  4. Add 100 μl of Standards or Samples into the respective wells.
  5. Cover the plate and incubate for 120 minutes at 37°C
  6. Aspirate and wash plate 4 times with Wash Buffer (1X) and blot residual buffer by firmly tapping plate upside down on absorbent paper. Wipe off any liquid from the bottom outside of the microtiter wells as any residue can interfere in the reading step.
  7. Add 100 μl of TMB Substrate in each well.
  8. Incubate the plate at 37°C for 30 minutes in dark. DO NOT SHAKE or else it may result in higher backgrounds and worse precision. Positive wells should turn bluish in color.
  9. Pipette out 100 μl of Stop Solution. Wells should turn from blue to yellow in color.Read the absorbance at 450 nm with a microplate reader.

Package Inserts

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