Certolizumab Antibody ELISA Kit

Catalog No. ABIN5684030

The Human Certolizumab Antibody ELISA Kit (ABIN5684030) is a Colorimetric ELISA Kit designed to quantify Human Certolizumab Antibody.

Quick Overview for Certolizumab Antibody ELISA Kit (ABIN5684030)

Target: Certolizumab Antibody

Reactivity: Human

Detection Method: Colorimetric

Method Type: Bridging ELISA

Application: ELISA

Sample Type: Plasma, Serum

Product Details

Purpose: Enzyme immunoassay for the semi-quantitative determination of free antibodies to Certolizumab pegol (CZP) in serum and plasma.

Analytical Method: Semi-Quantitative

Specificity: The solid phase (MTP) is coated by the drug Certolizumab Pegol. Due to the assay design this test only measures the free antibodies which are not bound to Certolizumab Pegol. Results are expressed in AU (arbitrary units).

Characteristics: The Antibody to Certolizumab pegol ELISA 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.

Components: Microtiter Plate
Negative Control
Positive Control
Dilution Buffer
Assay Buffer
Biotinylated CZP
Enzyme Conjugate
TMB Substrate Solution
Stop Solution
Wash Buffer, Concentrate (20x)
Adhesive Seal

Material not included: 1. Micropipettes (< 3 % CV) and tips to deliver 5-1000 μL.
2. Bidistilled or deionised water and calibrated glasswares (e.g. flasks or cylinders).
3. Wash bottle, automated or semi-automated microtiter plate washing system.
4. Microtiter plate reader capable of reading absorbance at 450 nm (reference wavelength at 600-650 nm is optional).
5. Absorbent paper towels, standard laboratory glass or plastic vials, and a timer.

Application Details

Application Notes: This kit has been developed for the detection of anti-drug antibodies in research and diagnostic uses. It is suitable for Therapeutical Drug Monitoring (TDM) purposes.

Comment:

The Anti Drug Antibody (ADA) ELISA kits are suitable also for being used by an automated ELISA processor.

Sample Volume: 10 μL

Plate: Pre-coated

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

Assay Precision: Intra-assay CV: <10%.
Inter-assay CV: <10%

Restrictions: For Research Use only

Handling

Storage: RT,4 °C

Storage Comment: The 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.

Expiry Date: 24 months

Target Details

Target: Certolizumab Antibody

Target Type: Anti-Drug Antibody

Background: The drug Certolizumab pegol (trade name Cimzia®) is a tumor necrosis factor alpha (TNFα) blocker and binds to human TNFα with a KD of 90pM. Certolizumab pegol is a recombinant, humanized antibody Fab' fragment, with specificity for TNFα, conjugated to an approximately 40  kDa polyethylene glycol (PEG). The Fab'fragment is manufactured in E. coli and is subsequently subjected to purification and conjugation to PEG2MAL40K, to generate Certolizumab pegol. Steady-state concentrations range from 0.5 to 90 μg/mL for a fixed dose of 400 mg of Certolizumab pegol. For patients developing anti-Certolizumab pegol antibodies, the steady state concentrations range from 0.5 to 75 μg/mL. Patients were tested at multiple time points for antibodies to Certolizumab pegol during studies CD1 and CD2. The overall percentage of antibody positive patients was reported to be 8 % in patients continuously exposed to Certolizumab pegol, of which approximately 80 % were neutralizing in vitro. In contrast to the common assumption that PEG is considered as a biologically inert material with no immunogenicity and antigenicity, both pre-existing and therapeutically induced anti-PEG antibodies have been found in the general population as well as patients receiving PEGylated therapeutics. In 1984, it was first observed that 0.2 % and 3.3 % of normal subjects and untreated allergy patients, respectively, exhibited relatively high titers of mostly anti-PEG IgM. The existence of anti-PEG immunity has brought complications to PEGylated protein therapeutics, such as the loss of therapeutic efficacy and increased adverse events. To manage these challenges, monitoring the presence of anti-PEG antibodies before and during PEGylated drug treatment might be beneficial.