Anti-Vedolizumab (Entyvio®) ADA Qualitative ELISA Kit

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SKU:
HUMB00019
€999

Description

Anti-Vedolizumab (Entyvio®) ADA Qualitative ELISA Kit

Enzyme-linked immunosorbent assay (ELISA) for the qualitative determination of specific antibodies to Vedolizumab (Entyvio®) in human serum and plasma. The ELISA Genie Antibody to Vedolizumab (Entyvio®) ELISA Kit is intended for the qualitative determination of Vedolizumab anti-drug antibodies in serum and plasma. It is for research use only.

Anti-Vedolizumab (Entyvio®) ADA Qualitative ELISA Kit test principle

The ELISA 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-Vedolizumab (Entyvio®) Product Information

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

Alternative Names

Anti-Human lymphocyte α4β7 integrin

Entyvio

Anti-Vedolizumab ADA Qualitative - Key Information

Vedolizumab (Entyvio®) mode of action

Vedolizumab is a recombinant humanized IgG1 monoclonal antibody directed against the human lymphocyte α4β7 integrin, a key mediator of gastrointestinal inflammation. Vedolizumab binds specifically to α4β7 integrin but does not bind to, or inhibit function of, α4β1 or αEβ7 integrins. Inhibition of the α4β7 integrin is a shared mechanism with natalizumab, however vedolizumab binds solely to the α4β7but not the α4β1 integrin, unlike natalizumab which binds to both.

As a result, natalizumab modulates the systemic immune system and is associated with other side effects such as progressive multifocal leukoencephalopathy (PML). Non-clinical studies have shown that the pharmacodynamic effects of vedolizumab are reversible upon removal of the antibody: pharmacologic activity of cells inhibited by vedolizumab could be partially restored within 24 hours after removal, with near complete restoration within 4 days. There are no known drug interactions as vedolizumab is a humanized antibody and does not modulate production of cytokines, which is known to affect drug metabolism

Vedolizumab (Entyvio®) uses

It is used in the treatment of moderate to severe active ulcerative colitis and Crohn's disease for patients who have had an inadequate response with, lost response to, or were intolerant to inhibitors of tumor necrosis factor alpha (TNF alpha) or other conventional therapies. By blocking its primary target, α4β7 integrin, vedolizumab reduces inflammation in the gut. Vedolizumab binds to α4β7 integrin, a key mediator of gastrointestinal inflammation expressed on the surfaces of T and B lymphocytes. By selectively inhibiting the α4β7 integrin, vedolizumab inhibits adhesion of lymphocytes to its natural ligand, mucosal addressin cell adhesion molecule-1 (MAdCAM-1), thereby preventing lymphocytic cells from entering the gut lamina propria and gut-associated lymphoid tissue (GALT). Specifically inhibiting this pathway alleviates GI inflammation without impairing systemic immune responses.

Vedolizumab is indicated for adult patients with moderately to severely active ulcerative colitis or Crohn's disease who have had an inadequate response with, lost response to, or were intolerant to a tumor necrosis factor (TNF) blocker or immunomodulator; or had an inadequate response with, were intolerant to, or demonstrated dependence on corticosteroids.

Vedolizumab (Entyvio®) immunogenicity  

As with any biologic therapy, immunogenicity, in the form of anti-vedolizumab antibodies (AVAs), occurs with following administration. The demonstration of anti-vedolizumab antibodies during treatment with vedolizumab (Entyvio®) is a major concern. Monitoring for the presence and/or quantitation of specific antibodies during clinical trials is an important issue for treatment regimen follow-up. The ELISA Genie Anti-Vedolizumab ADA Qualitative ELISA Kit can be efficiently used for monitoring vedolizumab anti-drug antibodies in biological samples and is for research use only.

Anti-Vedolizumab (Entyvio®) 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 vedolizumab

1 x 0.25 mL

Reactive Control
Ready-to-use. Contains vedolizumab-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 50 mL

Wash Buffer concentrate (20x)
Contains Buffer with Tween 20.

2 x 1

Adhesive Film
For covering of Microtiter Plate during incubation.


Anti-Vedolizumab (Entyvio®) ELISA 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 foil. Incubate 30 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 HRP-Conjugated Probe into each well.

6

Cover the plate with adhesive foil. Incubate 30 min at room temperature (18- 25°C).

7

Remove adhesive foil. 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.

Trademarks

Remicade® is a registered trademark of Janssen Biotech Inc.

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