PCSK9 Neutralization Rate / Evolocumab ELISA Assay Kit

$1,055.00

The PCSK9 Neutralization Rate / Evolocumab ELISA Assay Kit (RPE) is an enzyme immunoassay for the quantitative in-vitro determination of free PCSK9, the PCSK9 neutralization rate and the free antibody Evolocumab in serum samples. The PCSK9 Neutralization Rate / Evolocumab ELISA Assay Kit is for Research Use Only.

PCSK9 Neutralization Rate / Evolocumab ELISA Assay (RPE)

The PCSK9 Neutralization Rate / Evolocumab ELISA Assay is For Research Use Only

Size: 1x96wells
Sensitivity: Evolocumab: 0.2 µg/mL; PCSK9: 40 ng/ml
Dynamic Range: Evolocumab: 0.2 – 24 µg/mL; PCSK9 in sample without Evolocumab: 40-800 ng/ml; PCSK9 in sample with Evolocumab: 40-350 ng/ml
Incubation Time: Min 16 hours, Max 22 hours
Sample Type: Serum
Sample Size: 60 µL

Controls Included


Assay Background

Proprotein convertase subtilisin/kexin type 9 (PCSK9) has importance because it interferes with the lipoprotein in particular the lipoprotein particles (LDL) homeostasis. PCSK9 binds to the lipoprotein particles receptor (LDLR) which transports fat molecules within the extracellular fluid. After LDL binding to the LDL receptor the complex of LDLR/PCSK9/LDL gets degraded in the hepatocytes upon internalization. Importantly, if PCSK9 does not bind to the complex, the LDLR can return to the surface of the cell and can thereby continue to remove LDL-particles from the bloodstream.

Agents which block PCSK9 can lower LDL particle concentrations in the blood. The therapeutically monoclonal antibody against PCSK9 Evolocumab, was approved by the U.S. Food and Drug Administration in 2015 for lowering LDL-particle concentrations when statins and other drugs were not sufficiently effective or poorly tolerated.

To monitor the antibody (Evolocumab) concentration and the degree of neutralization of PCSK9 during Evolocumab therapy is beneficial to identify the ideal drug does for each patient individually. The PCSK9 Neutralization Rate / Evolocumab ELISA Assay Kit (RPE) allows the simultaneously determination of the free PCSK9 and the free Evolocumab in one ELISA assay.


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Additional Information

Assay Principle


The PCSK9 Neutralization Rate / Evolocumab ELISA Assay Kit (RPE) is performed in a 96-well microplate. The wells of the microplate are pre-coated with a specific capture antibody against human PCSK9 that binds free PCSK9 from the patient sample. A simultaneous incubation of the PCSK9 calibrators (with and without the addition of Evolocumab), the samples and the detection conjugate (Evolocumab peroxidase conjugate) is carried out. The incubation time is 16 to 22 hours at 2 to 8°C. After washing, the color substrate TMB (tetramethylbenzidine) is added to the wells. The enzymatic color reaction is stopped using a sulphuric acid solution. A color change from blue to yellow is optically detected.

Using a wavelength of 450 nm (reference value 620 nm), the optical density (OD) of the reaction product is measured with a suitable microplate reader. With the aid of the two calibrations, an evaluation procedure can be carried out to determine the concentrations of free PCSK9 and Evolocumab in the patient samples using the measured OD values. Determination of the therapeutic antibody is based on the principle that the presence of Evolocumab in patient samples leads to a systematic reduction in the recovery of PCSK9. The evaluation is performed using non-linear regression (Marquardt-Levenberg algorithm) of a logarithmic model from enzyme kinetics and the Langmuir isotherm from surface binding.

 

Documents

Product Manual


Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

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