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Quantification of Sorafenib in Human Serum by Competitive Enzyme-Linked Immunosorbent Assay
Tetsuya Saita1, Yuta Yamamoto, Satoshi Noda
1Applied Life Science Department, Faculty of Biotechnology and Life Science, Sojo University.
Abstract:
The multikinase inhibitor sorafenib has been used in the treatment of hepatocellular carcinoma, renal cell carcinoma, and differentiated thyroid carcinoma. Here we have demonstrated the production of the first specific antibody against sorafenib. Anti-sorafenib serum was obtained by immunizing mice with an antigen conjugated with bovine serum albumin and carboxylic modified 4-(4-aminophenoxy)-N-methyl-2-pyridinecarboxamide (AMPC) using the N-succinimidyl ester method. Enzyme labeling of sorafenib with horseradish peroxidase was similarly performed using carboxylic modified AMPC. A simple competitive enzyme-linked immunosorbent assay (ELISA) for sorafenib was developed using the principle of direct competition between sorafenib and the enzyme marker for anti-sorafenib antibody, which had been adsorbed by the plastic surface of a microtiter plate. Serum sorafenib concentrations lower than 0.04 µg/mL were reproducibly measurable using the ELISA. This ELISA was specific to sorafenib and showed very slight cross-reactivity (2.5%) with a major metabolite, sorafenib N-oxide. The values of serum sorafenib levels from 32 patients measured by this ELISA were comparable with those measured by HPLC, and there was a strong correlation between the values determined by the two methods (Y=1.016X-0.137, r=0.979). The specificity and sensitivity of the ELISA for sorafenib should provide a valuable new tool for use in therapeutic drug monitoring and pharmacokinetic studies of sorafenib.
Insights
Researchers developed a novel enzyme-linked immunosorbent assay (ELISA) for sorafenib, a multikinase inhibitor. This sensitive and specific ELISA allows for accurate therapeutic drug monitoring and pharmacokinetic studies of sorafenib in patients.
Area of Science:
- Biochemistry
- Immunology
- Pharmacology
Background:
- Sorafenib is a multikinase inhibitor used to treat hepatocellular carcinoma, renal cell carcinoma, and differentiated thyroid carcinoma.
- Accurate monitoring of sorafenib levels is crucial for effective therapeutic drug monitoring and pharmacokinetic studies.
Purpose of the Study:
- To develop and validate a specific antibody against sorafenib.
- To create a sensitive and specific competitive enzyme-linked immunosorbent assay (ELISA) for quantifying serum sorafenib concentrations.
Main Methods:
- Production of a specific anti-sorafenib antibody by immunizing mice with a sorafenib-conjugated antigen (AMPC-BSA).
- Development of a competitive ELISA using horseradish peroxidase-labeled sorafenib and anti-sorafenib antibody immobilized on a microtiter plate.
- Validation of the ELISA by comparing results with high-performance liquid chromatography (HPLC) in 32 patients.
Main Results:
- The developed ELISA can reproducibly measure serum sorafenib concentrations as low as 0.04 µg/mL.
- The assay demonstrated high specificity for sorafenib with minimal cross-reactivity (2.5%) with its metabolite, sorafenib N-oxide.
- Serum sorafenib levels measured by ELISA showed strong correlation (r=0.979) with HPLC measurements.
Conclusions:
- The novel anti-sorafenib ELISA is a sensitive, specific, and reliable tool for therapeutic drug monitoring.
- This assay will be valuable for pharmacokinetic studies and optimizing sorafenib treatment in patients.
- The developed immunoassay offers a practical alternative to existing methods for sorafenib quantification.
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