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New ELISA Method for Analysing Cetuximab

by Preethi Sivaswaamy Mohana on February 24, 2018 at 11:33 AM
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New ELISA Method for Analysing Cetuximab

Highly sensitive and selective ELISA can be used to determine cetuximab (CET) in human plasma samples, according to a new study. The findings of the study are published in the journal Current Pharmaceutical Analysis.

CET is an epidermal growth factor receptor inhibitor to treat cancers. It was captured by specific epidermal growth factor receptor (EGFR) antigen protein immobilized onto a 96-well assay plate.

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The CET- EGFR complex formed onto the plate wells were quantified, for the first time, using alkaline phosphatase enzyme labeled anti-human IgG (ALP-IgG) and para-nitrophenyl phosphate substrate (pNPP) as a chromogenic substrate for alkaline phosphatase enzyme. The optimum conditions for conducting the proposed ELISA were established and its analytical performance was evaluated as per the guidelines for the validation of immunoassays for bioanalysis of therapeutic monoclonal antibody. The assay limit of detection was 0.0015 μg/mL and the effective working dynamic range was 0.005-6.25 μg/mL. The accuracy and precision of the assay were proved.

The present assay with high throughput analysis very easy to perform in a 96-well plate and permits an operator to analyze a batch of ~ 200 samples, in triplicate. This facilitates the processing of a large number of samples in a clinical setting.
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The developed method eliminates the need for pretreatment of plasma samples by affinity chromatography or other sophisticated equipment. ELISA for the CET is expected to contribute in studying its PK, PD and TDM. The used reagents are commercially available, and the color product can be easily measured by inexpensive equipment available in biological laboratories.



Source: Eurekalert
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