Description

Datasheet

Golimumab (Simponi®) ELISA Kit

Enzyme-linked immunosorbent assay (ELISA) for the quantitative determination of free Golimumab (Simponi®) in serum and plasma. The Assay Genie Golimumab ELISA has been especially developed for the quantitative analysis of free Golimumab in serum and plasma samples.

Golimumab (Simponi®) ELISA Kit test principle

Solid phase enzyme-linked immunosorbent assay (ELISA) based on the sandwich principle. Standards and diluted samples (serum or plasma) are incubated in the micro-titre plate coated with the reactant for golimumab (Simponi®). After incubation, the wells are washed. A horse radish peroxidase (HRP) conjugated probe is added and binds to golimumab (Simponi®) captured by the reactant on the surface of the wells. Following incubation, wells are washed, and the bound enzymatic activity is detected by addition of chromogen-substrate. The colour developed is proportional to the amount of golimumab in the sample or standard. Results of samples can be determined directly using the standard curve.

Golimumab (Simponi®) Product Information

Information	Description
Application	Free drug
Required Volume (uL)	10
Total Time (min)	70
Sample Type	Serum, plazma and other biological fluids
Number of Assays	96
Detection Limit (ng/mL)	100 (ng/mL)
Spike Recovery (%)	85-115%
Shelf Life (year)	1
Alternative Names	Tumour Necrosis Factor alpha Simponi

Golimumab (Simponi®) - 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®) treatment

Golimumab was approved to be used in the underlined diseases alone or combination with methotrexate (MTX). According to American College of Rheumatology (ACR) criteria, golimumab 50 or 100 mg every 4 weeks in combination with MTX in methotrexate-naïve (GO-BEFORE) or -experienced (GO-FORWARD) rheumatoid arthritis patients was more effective than MTX alone to overcome the symptoms at week 14 and/or 24.

In rheumatoid arthritis treated with other anti-TNF agents (GO-AFTER) before, golimumab 50 or 100 mg every 4 weeks was more effective than placebo at week 14 and/or 24. These affirmative influences were also shown for psoriatic arthritis GO-REVEAL and ankylosing spondylitis GO-RAISE studies.

Golimumab (Simponi®) 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	Golimumab Standards A-E, High Level Control, Low Level Control 3000; 1000; 300; 100; 0 ng/mL Ready to use. Used for construction of the standard curve. Contains golimumab (Simponi®), human serum, stabilizer 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	Peroxidase Conjugate Red coloured. Ready to use. Contains peroxidase (POD) conjugate, stabilizer and <0,1% NaN3
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.

Golimumab (Simponi®) ELISA Protocol

Steps	Protocol
1	Pipette 100µl of Assay Buffer non-exceptionally into each of the wells to be used.
2	Pipette 10 µL of each ready-to use Standards, High Level Control, Low Level Control and 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)
3	Cover the plate with adhesive film. Briefly mix contents by gently shaking the plate. 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 10 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.