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Lookup NU author(s): Dr David Jamieson,
Dr Nicola Cresti,
Dr Mark Verrill,
Professor Alan Boddy
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Trastuzumab is a therapeutic monoclonal antibody against the Her2 oncoprotein, which is over-expressed in approximately 30% of breast cancers, and is now used routinely in the management of early and metastatic Her2+ disease. However, not all Her2+ breast cancer patients respond to trastuzumab and the pharmacodynamic and pharmacokinetic parameters behind this variation in response are unknown. Pharmacological investigations into variable response to trastuzumab have been hampered by the lack of a published feasible method to determine trastuzumab concentration in plasma. Here we describe the development and validation of a cell-based ELISA to measure trastuzumab in human plasma. The assay specifically measures the interaction between trastuzumab and Her2 and has a dynamic range of between 10 and 120 microg/ml. The mean intra-assay and inter-assay variability of the ELISA was 9%. Trastuzumab in plasma was stable for at least 10 weeks at -20 degrees C and 72 h at 4 degrees C, and was unaffected by 5 freeze/thaw cycles. Having validated the assay, the trough plasma trastuzumab concentrations of 30 patients being treated for metastatic or early disease were measured. The median trough concentration was 62 (range 21 to 441) microg/ml. This cell-based ELISA method has undergone appropriate validation and is suitable for quantification of trastuzumab in the plasma of patients treated with Herceptin.
Author(s): Jamieson D, Cresti N, Verrill MW, Boddy AV
Publication type: Article
Publication status: Published
Journal: Journal of Immunological Methods
ISSN (print): 0022-1759
ISSN (electronic): 1872-7905
Publisher: Elsevier BV
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