Description

Datasheet

Anti-Infliximab ADA Qualitative ELISA Kit

Enzyme-linked immunosorbent assay for the quantitative determination of antibodies to infliximab Remsima® in serum and plasma. Remsima® the world's first biosimilar mAb (approved in 2013 by EMA). The Committee for Medicinal Products for Human Use (CHMP) decided that, in accordance with EU requirements, Remsima® has been shown to have a comparable quality, safety and efficacy profile to Remicade. Infliximab (Remsima®) has been associated with the development of anti-infliximab antibodies, even some were reported to be neutralizing, in various percentages of patients during therapy with the drug Remsima®. This could lead to severe complications.

The Assay Genie Antibody to Infliximab® ELISA Kit can be efficiently used for monitoring infliximab anti-drug antibodies (ADA) in biological samples and is for research use only.

Anti-Infliximab (Remsima®) ADA Qualitative ELISA Kit test principle

The Assay Genie Antibody to infliximab biosimilar (Remsima®) ELISA is a sandwich assay for the determination of antibodies against infliximab biosimilar in serum and plasma samples. During the first incubation period, antibodies to infliximab biosimilar in patient serum/plasma samples are captured by the drug infliximab biosimilar (Remsima®) coated on the wall of the microtiter wells. After washing away the unbound components from samples, a peroxidase-labelled specific 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 infliximab in the sample.

Anti-Infliximab 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 Remsima

Anti-Infliximab (Remsima®) ADA Qualitative - Key Information

Infliximab mode of action

Infliximab (Remsima®) is a chimeric monoclonal antibody and used to treat autoimmune disorders. Infliximab reduces the amount of active human tumour necrosis factor alpha (hTNF alpha) in the body by binding to it and preventing it from signalling the receptors for TNF alpha on the surface of various cell types. TNF alpha is one of the key cytokines that triggers and sustains the inflammatory reactions.

Infliximab uses

Remsima, is the world’s first biosimilar mAb (approved in 2013 by EMA). The Committee for Medicinal Products for Human Use (CHMP) decided that, in accordance with EU requirements, Remsima has been shown to have a comparable quality, safety and efficacy profile to Remicade. Remsima is a tumor necrosis factor alpha (TNF-alpha) antagonist used to treat rheumatoid arthritis, ankylosing spondylitis, ulcerative colitis, adult Crohn's disease, plaque psoriasis, and psoriatic arthritis.

Infliximab immunogenicity

One of the major concern, despite of its wide usage, is potential development of anti-infliximab antibodies (ATI) which in turn may interfere with infliximab (Remsima®) efficacy as mainly judged by observing the relapse of signs and symptoms of disease and necessitate dose-escalation or potentially ending up the treatment. In this context, demonstration of anti-infliximab antibodies during treatment with infliximab (Remsima®) has a major concern and monitoring for the presence and/or quantitation of specific antibodies during clinical trials is an important issue for follow up of the treatment regimens.

The Assay Genie® ATI ELISA Kit can be efficiently used for monitoring infliximab-specific antibodies during therapy and is for research use only. With this Assay Genie® ELISA test, anti-infliximab antibodies can be detected and is for research use only.

Anti-Infliximab (Remsima®) ADA Qualitative ELISA Kit Contents

Size	Kit Contents
1x12x8	Microtiter Plate Break apart strips. Microtiter plate with 12 rows each of 8 wells coated with infliximab biosimilar.
1x0.25	Reactive Control Ready-to-use. Contains infliximab-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 12 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 50mL	Wash Buffer, concentrate (20x) Contains Buffer with Tween 20.
2x1	Adhesive Film For covering of Microtiter Plate during incubation.

Anti-Infliximab (Remsima®) ADA Qualitative ELISA Kit Protocol

Steps	Protocol
1	Pipette 100µl of Assay Buffer non-exceptionally into each of the wells to be used.
2	QUALITATIVE ELISA TEST FORMAT Pipette 10 µL of ready-to use Negative Control, Reactive Control, and Samples into the respective wells of microtiter plate. Wells A1: Negative Control B1: Negative Control C1: Reactive Control D1 and on: Sample (Serum/Plasma)
3	Cover the plate with adhesive film. Briefly mix contents by gently shaking the plate. Incubate 60 min at room temperature (18-25°C).
4	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.
5	Pipette 100 µL of ready-to use Peroxidase Conjugate into each well.
6	Cover the plate with adhesive film. 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 20 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.