Nivolumab Specific ELISA Kit

Catalog No. ABIN5012827

This Colorimetric ELISA kit is designed for the quantitative measurement of Human Nivolumab Specific.

Quick Overview for Nivolumab Specific ELISA Kit (ABIN5012827)

Target: Nivolumab Specific

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Plasma, Serum

Product Details

Purpose: Enzyme immunoassay for the specific quantitative determination of free Nivolumab in serum and plasma

Analytical Method: Quantitative

Specificity: There is no cross reaction with any other proteins present in native human serum. A screening test was performed with 48 different native human sera. All produced OD450/620 nm values (ranged from 0.043 to 0.100) less than the mean OD (0.150) of standard D (20 ng/mL). In addition, binding of Nivolumab to the solid phase is inhibited by recombinant human PD-1 protein in a concentration dependent manner. Therefore, the Nivolumab ELISA (mAb-Based) measures the biologically active free form of Nivolumab, i.e. not pre-occupied by human PD-1 antigen. No cross reaction was observed with sera spiked with the other therapeutic antibodies including Cetuximab, Aflibercept, Rituximab, Trastuzumab, Bevacizumab, Vedolizumab, Infliximab and Adalimumab at concentrations up to 200 μg/mL. All produced mean OD450/620 nm values (ranged from 0.024 to 0.030) less than standard D.

Components: plate, standards, assay buffer, conjugate, TMB, HCl, wash buffer

Material not included: normal lab equipment for performing ELISA assays

Application Details

Sample Volume: 20 μL

Assay Time: 1.5 h

Plate: Pre-coated

Protocol: This ELISA is based on Nivolumab-specific mouse monoclonal antibody (catcher Ab, clone NY-11C3). Diluted standards and samples are incubated in the microtiter plate coated with IG-NY-11C3 mAb. After incubation, the wells are washed. A horseradish peroxidase (HRP)-conjugated anti-human IgG antibody is added and binds to the Fc part of Nivolumab. Following incubation, wells are washed and the bound enzymatic activity is detected by addition of chromogen-substrate. The color developed is proportional to the amount of Nivolumab in the sample or standard. Results of samples can be determined by using the standard curve.

Reagent Preparation:

Just the wash buffer has to be prepared by diluting the stock solution 1:20. All other reagents are ready to use.

Sample Collection: normal serum or plasma collection

Sample Preparation:

dilute the samples 1:20 with assay buffer

Calculation of Results:

The results are read from a standard curve.

Assay Precision: < 10%

Restrictions: For Research Use only

Handling

Preservative: Sodium azide

Storage: 4 °C

Expiry Date: 24 months

Target Details

Target: Nivolumab Specific

Background: The drug Nivolumab (Opdivo®) is a fully human IgG4 monoclonal antibody that binds specifically to programmed death-1 (PD-1), a negative regulatory receptor expressed by activated T and B lymphocytes. Binding of Nivolumab to the PD-1 receptor blocks its interaction with the ligands, PDL1 and PD-L2, thereby attenuating PD-1-mediated inhibition of the immune response, including the anti-tumor immune response. According to the prescribing information, Nivolumab at 3 mg/kg every 2 weeks has been shown to be safe and efficacious in the treatment of advanced melanoma, refractory non-small cell lung cancer (NSCLC), and advanced renal cell carcinoma, and it is being studied in patients with other solid tumors and hematologic malignancies. Following the recommended dose regimen (3 mg/kg every 2 week), the mean Cmin (trough) and Cmax concentrations of Nivolumab were reported to be 15 μg/mL and 68.8 μg/mL, respectively. Identification of biomarkers for (non-) response and risk factors for adverse drug reactions that might be related to serum drug levels and maintaining the effective concentration in order to potentially avoid some side effects with a reliable method might be beneficial.