Anti-Infliximab (Remsima®) ADA Quantitative ELISA Kit

(No reviews yet) Write a Review
SKU:
HUMB00007
€899

Description

Anti-Infliximab ADA Quantitative ELISA Kit

Enzyme-linked immunosorbent assay for the quantitative determination of antibodies to infliximab Remsima® in serum and plasma. Remsima® is the world's first biosimilar mAb (approved in 2013 by EMA). The Agencys 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®) was associated to 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 ELISA Genie Antibody to Infliximab® ELISA Kit can be used to monitor Infliximab anti-drug antibodies (ADA) and is for research use only.

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

The ELISA Genie Antibody to infliximab biosimilar (CT-P13) (Remsima®) ELISA is a sandwich assay for the determination of antibodies against infliximab biosimilar (CTP13) in serum and plasma samples. During the first incubation period, antibodies to infliximab biosimilar (CT-P13) in patient serum/plasma samples are captured by the drug infliximab biosimilar (CT-P13) (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 anti-infliximab antibodies in the sample.

Anti-Infliximab (Remsima®) ADA Quantitative ELISA Product Information

Information Description
Application
Free drug
Required Volume (μl)
10
Total Time (min)
140
Sample Type
Serum, Plasma
Number of Assays
96
Detection Limit (ng/mL)
31 (ng/mL)
Spike Recovery (%)
85-115%
Shelf Life (year)
1

Alternative Names

Tumour Necrosis Factor Alpha

Remsima

Anti-Infliximab (Remsima®) ADA Quantitative - 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, 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 presented comparable quality, safety and efficacy profile to Remicade. Infliximab (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 concerns, despite its wide usage, is the potential development of anti-infliximab antibodies which in turn may interfere with infliximab (Remsima®) efficacy. This was determined based on the return of signs and symptoms of disease. This issue could lead to a dose-escalation of the therapeutic or a potential termination of treatment. In this context, the presence of anti-infliximab antibodies during treatment with infliximab (Remsima®) is a major concern and monitoring for the presence and/or quantitation of specific antibodies during clinical trials is often a necessity for follow up of the treatment regimens.

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

Anti-Infliximab ADA 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 biosimilar

7 x 1 mL

AIR 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 biosimilar, human serum 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

Confirmation Reagent
Ready to use. Contains optimized concentration of the infiliximab
biosimilar, proteins, stabilizer 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.

2 x 1

Semi-Log Graph Paper
For constructing standard curve and calculation of results.

Anti-Infliximab ADA Quantitative ELISA Protocol

Steps Protocol

1

QUANTITATIVE ELISA TEST FORMAT
Pipette 100 µL of ready-to use Standards, High Level Control, Low Level Control, Diluted Samples into the respective wells of microtiter plate

Wells:

A1: Negative Control
B1: Negative Control
C1: Reactive Control
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.

Trademarks

REMSIMA is a trademark of CELLTRION, INC.

View AllClose