Anti-Omalizumab (Xolair®) ADA Qualitative ELISA Kit

(No reviews yet) Write a Review
SKU:
HUMB00040
€899

Description

Anti-Omalizumab (Xolair®) ADA Qualitative ELISA Kit

Enzyme-linked immunosorbent assay (ELISA) for the qualitative determination (screening) of antibodies to Omalizumab (Xolair®) in serum and plasma. The ELISA Genie Antibody to omalizumab (Xolair®) ELISA Kit is intended for the qualitative determination of antibodies to omalizumab (Xolair®) in serum and plasma. It is for professional use only.

Anti-Omalizumab (Xolair®) ADA Qualitative ELISA Kit test principle

The Elisa Genie Antibody to omalizumab (Xolair®) ELISA is a sandwich assay for the determination of antibodies against omalizumab in serum and plasma samples. During the first incubation period, antibodies to omalizumab (AT0) in patient serum/ plasma samples are captured by the drug omalizumab (Xolair®) 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 ATO in sample.

Anti-Omalizumab (Xolair®) ADA Qualitative 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)
plus/minus
Spike Recovery (%)
-
Shelf Life (year)
1

Alternative Names

IgG1k mAb

Xolair

Anti-Omalizumab (Xolair®) ADA Qualitative - Key Information

Omalizumab (Xolair®) mode of action

Omalizumab is a recombinant DNA-derived humanized IgG1k monoclonal antibody that selectively binds to human immunoglobulin E (IgE). Omalizumab (Xolair) is produced by a Chinese hamster ovary cell suspension culture in a nutrient medium containing the antibiotic gentamicin. Omalizumab (Xolair) inhibits the binding of IgE to the high-affinity IgE receptor (FceRI) on the surface of mast cells and basophils. Reduction in surface-bound IgE on FcεRI-bearing cells limits the degree of release of mediators of the allergic response. Omalizumab does not bind to cell surface IgE, so it does not directly activate mast cells or basophils. The decrease in free IgE levels results in a reduced amount of FcεRI receptors on mast cells, basophils, and antigen-presenting cells.

Omalizumab (Xolair®) uses

Xolair is used to treat severe, persisten asthma. Xolair binds to IgE (a class of antibodies normally secreted in allergic responses), which prevents their binding to mast cells and basophils. Most likely removed by opsonization via the reticuloendothelial system. Liver elimination of IgG includes degradation in the liver reticuloendothelial system (RES) and endothelial cells. Intact IgG is also excreted in bile. Liver elimination of IgG includes degradation in the liver reticuloendothelial system (RES) and endothelial cells. Intact IgG is also excreted in bile.

Omalizumab (Xolair®) immunogenicity

As with any biologic therapeutic, immunogenicity, in the form of anti-omalizumab antibodies can occur. The demonstration of anti-omalizumab antibodies during treatment with Omalizumab (Xolair®) is a major concern. The ELISA Genie Anti-Omalizumab (Xolair®) ADA Quantitative ELISA Kit can be efficiently used for monitoring anti-omalizumab antibodies in biological samples and is for research use only.

Anti-Omalizumab (Xolair®) 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 denosumab-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 Foil
For covering of Microtiter Plate during incubation.


Anti-Omalizumab (Xolair®) ADA Qualitative 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 film. Briefly mix contents by gently shaking the plate. 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 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

Xolair® is a trademark of Novartis AG.

View AllClose