Description

Datasheet

Anti-Infliximab (Remicade®) Quantitative ELISA Kit

Enzyme-linked immunosorbent assay (ELISA) for the quantitative determination of specific antibodies to infliximab in human serum and plasma with confirmation. Infliximab (Remicade®) was associated with the development of anti-Infliximab antibodies, with some reported to be neutralizing. The Assay Genie Antibody to Infliximab ELISA Kit can be efficiently used for monitoring Infliximab anti-drug antibodies (ADA) .

Anti-Infliximab Quantitative ELISA Kit test principle

The Assay Genie Antibody to infliximab (Remicade®) ELISA is a sandwich assay for the determination of antibodies against infliximab in serum and plasma samples. During the first incubation period, antibodies to infliximab (ATI) in patient serum/ plasma samples are captured by the drug infliximab (Remicade®) 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 ATI in sample.

Anti-Infliximab Quantitative 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)	15 (ng/mL)
Spike Recovery (%)	85-115%
Shelf Life (year)	1
Alternative Names	Tumour Necrosis Factor Alpha Remicade

Anti-Infliximab Quantitative ELISA - Key Information

Infliximab mode of action

Infliximab (Remicade®) 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 treatment

Infliximab (Remicade®) is used for the treatment of psoriasis, Crohn’s disease, ankylosing spondylitis, psoriatic arthritis, rheumatoid arthritis, ulcerative colitis, and is approved by the FDA.

Infliximab immunogenicity

One of the major concerns, despite its wide usage, is the potential development of anti-infliximab antibodies (ATI) which in turn may interfere with infliximab (Remicade®) 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 (Remicade®) 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 anti-drug antibodies (ADA) and is for research use only.

Anti-Infliximab Quantitative 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
7 x 0.3 mL	Infliximab Standards A-E, High Level Control, Low Level Control 500; 250; 125; 62; 0 ng/mL Ready to use. Used for construction of the standard curve. Contains antibody to infliximab, human serum and <0,1% NaN3
1 x 50 mL	Assay Buffer Blue coloured. Ready to use. Contains proteins, RF blockers 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-Infliximab Quantitative ELISA Kit Protocol

Steps	Kit content
1	QUANTI QUANTITATIVE ELISA TEST FORMAT Pipette 100 µL of ready-to use Standards, High Level Control, Low Level Control, Low Level and Pre-diluted Samples into the respective wells of microtiter plate. Wells: A1: Standard A B1:Standard B C1: Standard C D1: Standard D E1: Standard E F1: High Level Control G1: Low Level Control H1 and on : Sample (Serum / Plasma)
2	Cover the plate with adhesive film. Briefly mix contents by gently shaking the plate. Incubate 60 min at room temperature (18-25°C).
3	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.
4	Pipette 100 µL of ready-to use Peroxidase Conjugate into each well.
5	Cover the plate with adhesive film. Incubate 60 min at room temperature (18- 25°C).
6	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.
7	Pipette 100 µL of TMB Substrate Solution into each well.
8	Incubate 20 min (without adhesive foil) at room temperature (18-25°C) in the dark
9	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.
10	Measure optical density with a photometer at 450/650 nm within 30 min after pipetting of the Stop Solution.