Infliximab (Remicade®)

The drug Infliximab (trade name Remicade®, Remsima®, Inflectra®) is a chimeric monoclonal antibody and used to treat auto-immune disorders. It reduces the amount of active tumor necrosis factor alpha (hTNFα) by binding to it and preventing it from signaling the receptors for TNFα on the surface of various cell types. TNFα is one of the key cytokines that triggers and sustains the inflammatory reactions. The drug is used for the treatment of psoriasis, Crohn's disease, ankylosing spondylitis, psoriatic arthritis, rheumatoid arthritis, ulcerative colitis. Single intravenous (IV) infusion of 3 mg/kg to 20 mg/kg showed a linear relationship between the dose administered and the maximum serum concentration. The volume of distribution at steady state was independent of dose and indicated that Infliximab was distributed primarily within the vascular compartment. Identification of biomarkers for (non-)response and risk factors for adverse drug reactions that might be related to serum concentrations and maintaining the effective concentration of Infliximab in order to potentially avoid some side effects with a reliable method might be beneficial. Enzyme immunoassay (ELISA) test kit for the determination of free Infliximab (Remicade®, Remsima®) in serum and plasma. The catcher is TNF-α and the detection antibody is a HRP-labeled mAb binding at the Fc part of the drug. Due to the assay design there is full cross reactivity with other TNF-α catchers. This kit is for research use only, not for use in diagnostic procedures.
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Protocol PDF Click for Protocol
Catalog Number TK01451
Species Human
Design Enzyme immunoassay (ELISA) technique.
Standards 5 standards, ready to use.
Controls None provided.
Sample Types Serum and plasma.
Sample Volume 10 µL
Assay Desc. 60 min. incubation (RT) + 30 min. (RT) + 15 min. (RT) = 1 hour, 45 min. total incubation time.
Standard Range 0 / 6 - 200 ng/mL
Sensitivity 5 ng/mL
Other Names
Storage 2 - 8°C
Postscript For research use only, not for use in diagnostic procedures.

Intended use

This ELISA kit applies to the in vitro quantitative determination of Infliximab (Remicade®) concentrations in serum, plasma and other biological fluids.

Test principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antigen specific to Infliximab (Remicade®). Standards or samples are added to the micro ELISA plate wells and combined with the specific antigen. Then a biotinylated detection antigen specific for Infliximab (Remicade®) and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Infliximab (Remicade®), biotinylated detection antigen and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Infliximab (Remicade®). You can calculate the concentration of Infliximab (Remicade®) in the samples by comparing the OD of the samples to the standard curve.

Typical data

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.


Intra-assay Precision (Precision within an assay):
3 samples with low, middle,high concentration of Infliximab (Remicade®). And the sample of Infliximab (Remicade®) were tested 30 times on one plate, respectively.
Inter-assay Precision (Precision between assays):
3 samples with low, middle,high concentration of Infliximab (Remicade®). And the sample of Infliximab (Remicade®) were tested on 3 different plates, 30 replicates in each plate.


Matrices listed below were spiked with certain level of Infliximab (Remicade®) and the recovery rates were calculated by comparing the measured value to the expected amount of Infliximab (Remicade®) in samples.

Matrix Recovery range (%) Average Recovery (%)
Serum (n=5) 88-99 98
EDTA plasma (n=5) 89-103 96
heparin plasma (n=5) 87-102 93


The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Infliximab (Remicade®) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

    Serum (n=5) EDTA plasma (n=5) heparin plasma (n=5)
1:2 Range (%) 89-102 88-100 90-103
Average (%) 94 95 95
1:4 Range (%) 87-104 91-100 93-107
Average (%) 95 93 95
1:8 Range (%) 90-103 90-101 92-103
Average (%) 93 93 93
1:16 Range (%) 94-106 88-99 87-105
Average (%) 93 93 93


The stability of Infliximab (Remicade®) is determined by the loss rate of activity. The loss rate of Infliximab (Remicade®) is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Item Specifications Storage
Micro ELISA Plate(Dismountable) 8 wells ×12 strips -20℃, 6 months
Reference Standard 2 vials
Concentrated Biotinylated Detection Ab (100×) 1 vial, 120 μL
Concentrated HRP Conjugate (100×) 1 vial, 120 μL -20℃(shading light), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 4℃, 6 months
Biotinylated Detection Ab Diluent 1 vial, 14 mL
HRP Conjugate Diluent 1 vial, 14 mL
Concentrated Wash Buffer (25×) 1 vial, 30 mL
Substrate Reagent 1 vial, 10 mL 4℃(shading light)
Stop Solution 1 vial, 10 mL 4℃
Plate Sealer 5 pieces
Product Description 1 copy
Certificate of Analysis 1 copy

Other supplies required

Microplate reader with 450 nm wavelength filter
High-precision transfer pipette, EP tubes and disposable pipette tips
Incubator capable of maintaining 37℃
Deionized or distilled water
Absorbent paper
Loading slot for Wash Buffer

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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