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Anti-Golimumab (Simponi®)ADA Qualitative ELISA Kit

SKU:
HUMB00016
Product Type:
ELISA Kit
ELISA Type:
Biosimilar ELISA
Biosimilar ELISA Type:
Qualitative
Applications:
ELISA
Reactivity:
Human
Analytes:
Golimumab (Simponi®)
Research Area:
Anti-TNF Alpha
€1,019
Frequently bought together:

Description

system_update_altDatasheet

Anti-Golimumab (Simponi®) ADA Qualitative ELISA Kit

Enzyme-linked immunosorbent assay (ELISA) for the qualitative determination of antibodies to Golimumab in serum and plasma. Golimumab (Simponi®) was associated to the development of anti-Golimumab antibodies, with some reported to be neutralizing in patients. The Assay Genie Anti-Golimumab ADA Qualitative ELISA Kit can be efficiently used for monitoring Golimumab anti-drug antibodies (ADA) and is for research use only.

Anti-Golimumab (Simponi®) ADA Qualitative ELISA Kit test principle

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

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

Alternative Names

Tumour Necrosis Factor alpha

Simponi

Anti-Golimumab ADA Qualitative - Key Information

Golimumab (Simponi®) mode of action

Golimumab (Simponi, CNTO-148) is a human immunoglobulin G1 kappa monoclonal antibody which is specific for proinflammatory cytokine, tumor necrosis factor- alpha (TNF alpha). TNF is a naturally occurring cytokine that is involved in normal inflammatory and immune responses. Golimumab binds to both the soluble and transmembrane bioactive forms of human TNF and prevent TNF from binding to its receptors and finally inhibits biological activity of TNF.

Golimumab (Simponi®) uses

In 2009, it was approved by FDA for the treatment of rheumatoid arthritis, psoriatic arthritis and ankylosing spondylitis in adult patients. Elevated levels of TNF are found in the synovial fluid of rheumatoid arthritis, including juvenile idiopathic arthritis, psoriatic arthritis, and ankylosing spondylitis patients and play an important role in both the pathologic inflammation and the joint destruction that are hallmarks of these diseases. Increased levels of TNF are also found in psoriasis (Ps) plaques. Its affinity for TNF in surface plasmon resonance assay was 17 pmol/L.

Golimumab (Simponi®) immunogenicity

As with any biologic therapeutic, immunogenicity, in the form of anti-golimumab antibodies, occurs with following administration. The demonstration of anti-golimumab antibodies during treatment with golimumab (Simponi®) 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 Assay Genie Anti-Golimumab ADA Qualitative ELISA Kit can be efficiently used for monitoring golimumab-specific antibodies in biological samples and is for research use only.

Anti-Golimumab ADA Qualitative ELISA Kit Contents

Size Kit Content

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 golimumab -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-Golimumab 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

Simponi is a registered trademark of JOHNSON & JOHNSON