Description

Datasheet

Anti-Adalimumab (Humira®) ADA Qualitative ELISA Kit

Enzyme-linked immunosorbent assay for the qualitative determination of antibodies to adalimumab in serum and plasma with confirmation. Adalimumab (Humira®) was associated to the development of anti-Adalimumab antibodies, with some reported to be neutralizing in patients. The Assay Genie Antibody to Adalimumab ELISA Kit can be efficiently used for monitoring Adalimumab anti-drug antibodies (ADA) in biological samples and is for research use only.

Anti-Adalimumab (Humira®) ADA Qualitative ELISA Kit test principle

The Assay Genie Antibody to Adalimumab (Humira®) ELISA is a sandwich assay for the determination of antibodies against adalimumab in serum and plasma samples. During the first incubation period, the drug adalimumab coated on the wall of the microtiter wells captures the antibodies to adalimumab in patient serum and plasma samples. After washing away the unbound components from samples, a Peroxidase-labelled conjugate is added to each well and then incubated. After a second washing step, the bound enzymatic activity is detected by addition of tetramethylbenzidine (TMB) chromogen-substrate. Finally, the reaction is terminated with an acidic stop solution. The intensity of the reaction colour is directly proportional to the concentration of antibodies to Adlimumab in sample.

Anti-Adalimumab (Humira®) ADA Qualitative ELISA Product Information

Information	Description
Application	Free drug
Required Volume (uL)	10
Total Time (min)	140
Sample Type	Serum, Plasma
Number of Assays	96
Detection Limit (ng/mL)	30 (ng/mL)
Spike Recovery (%)	85-115%
Shelf Life (year)	1
Alternative Names	Tumour Necrosis Factor Alpha Humira

Anti-Adalimumab (Humira®) ADA Qualitative - Key Information

Adalimumab mode of action

Adalimumab (Humira®) is a recombinant human IgG1 monoclonal antibody specific for human tumor necrosis factor alpha (TNF-a). Adalimumab (Humira®) was created using phage display technology resulting in an antibody with human derived heavy and light chain variable regions and human IgG1:k constant regions. Adalimumab (Humira®) is produced by recombinant DNA technology in a mammalian cell expression system and is purified by a process that includes specific viral inactivation and removal steps.

Adalimumab (Humira®) binds specifically to (TNF-a) and blocks its interaction with the p55 and p75 cell surface TNF receptors. Adalimumab (Humira®) also lyses surface TNF expressing cells in vitro in the presence of complement. Adalimumab (Humira®) does not bind or inactivate lymphotoxin (TNF-beta). TNF is a naturally occurring cytokine that is involved in normal inflammatory and immune responses. Elevated levels of TNF are found in the synovial fluid of rheumatoid arthritis, including juvenile idiopathic arthritis, psoriatic arthritis, and ankylosing spondylitis patients and play an important role in both the pathologic inflammation and the joint destruction that are hallmarks of these diseases. Increased levels of TNF are also found in psoriasis (Ps) plaques.

Adalimumab uses

Adalimumab is to treat various autoimmune diseases and works to treat inflammation. Adalimumab is administered to minimize pain and swelling due to various types of arthritis, including rheumatoid arthritis, psoriatic arthritis, juvenile idiopathic arthritis and ankylosing spondylitis. Adalimumab also treats various skin conditions including plaque psoriasis and hidradenitis suppurativa.

Adalimumab immunogenicity

When administered to patients, all therapeutic proteins have the potential to induce an unwanted immune response for example, to stimulate the formation of antidrug antibodies (ADAs). The impact of immune responses can range from no apparent effect to changes in pharmacokinetics, loss of effect and serious adverse events.

According to the manufacturer's product insert, the use of Adalimumab (Humira®) was associated to the development of anti-adalimumab antibodies, even some were reported to be neutralizing in various percentages of patients, during therapy with the drug Humira®. This has the possibility to lead to severe complications. The Assay Genie Antibody to Adalimumab ELISA Kit can be efficiently used for monitoring Adalimumab anti-drug antibodies (ADAs) in biological samples and is for research use only.

Anti-Adalimumab (Humira®) ADA Qualitative ELISA Kit Contents

Size	Kit Contents
1 x 12 x 8	Microtiter Plate Break apart strips. Microtiter plate with 12 rows each of 8 wells coated with reactant
1 x 0.25 mL	Reactive Control Ready-to-use. Contains adalimumab-reactive reagent, human serum, stabilizers and <0.1% NaN3.
1 x 0.5 mL	Negative Control Ready-to-use. Contains human serum, stabilizers and <0.1% NaN3
1 x 50 mL	Assay Buffer Blue coloured. Ready to use. Contains proteins and <0.1% NaN3.
1 x 12 mL	Peroxidase-Conjugate Red coloured. Ready to use. Contains peroxidase (POD) conjugate, stabilizer and preservatives.
1 x 12 mL	TMB Substrate Solution Ready to use. Contains TMB
1 x 12 mL	TMB Stop Solution Ready to use. 1N HCl
1 x 50 mL	Wash Buffer concentrate (20x) Contains Buffer with Tween 20.
2 x 1	Adhesive Film For covering of Microtiter Plate during incubation.

Anti-Adalimumab (Humira®) ADA Qualitative ELISA Kit Contents

Steps	Protocol
1	Pipette 100µl of Assay Buffer non-exceptionally into each of the wells to be used.
2	Pipette 10 µL of each ready-to use Standards, High Level Control, Low Level Control and Diluted Samples into the respective wells of microtiter plate. Wells A1: Standard A B1: Standard B C1: Standard C D1 and so on: Sample (Serum / Plasma)
3	Cover the plate with adhesive foil. Incubate 60 min at room temperature (18- 25°C).
4	Remove adhesive foil. Discard incubation solution. Wash plate 3 times each with 300µL of diluted Wash Buffer. Remove excess solution by tapping the inverted plate on a paper towel.
5	Pipette 100 µL of ready-to use Peroxidase Conjugate into each well.
6	Cover the plate with adhesive foil. Incubate 60 min at room temperature (18- 25°C).
7	Remove adhesive film. Discard incubation solution. Wash plate 3 times each with 300 µL of diluted Wash Buffer. Remove excess solution by tapping the inverted plate on a paper towel.
8	Pipette 100 µL of TMB Substrate Solution into each well.
9	Incubate 10 min (without adhesive foil) at room temperature (18-25°C) in the dark
10	Stop the substrate reaction by adding 100 µL of Stop Solution into each well. Briefly mix contents by gently shaking the plate. Colour changes from blue to yellow.
11	Measure optical density with a photometer at 450/650 nm within 30 min after pipetting of the Stop Solution.