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A label-free and reagent-free peptide mimotope capacitive biosensor has been developed for cancer drug (trastuzumab) quantification based on nonfaradic readout. The low sensitivity issue of capacitive biosensors was overcome with two innovations: peptide mimotope mixed self-assembled monolayer (SAM) biointerface and dilution of the analysis buffer. Signal amplification was achieved through dilution of phosphate-buffered saline (PBS) to tune Cdl to dominate the overall capacitance change upon target binding, which contribution is often negligible without dilution. After 1000× dilution, the limit of detection was lowered 500-fold (0.22 μg/mL) and the sensitivity was increased 20-fold [0.04192 (μg/mL)−1] in comparison with undiluted PBS. The proposed signal amplification strategy is more straightforward and practical compared to biorecognition element engineering and other strategies. The proposed method was further applied to planar electrodes for optimizing sensing response time to less than 1 min.

Therapeutic antibodies are antigenically similar to human antibodies and are difficult to detect in assays of human serum samples without the use of the therapeutic antibody's complementary antigen. Herein for the first time, we established a platform to detect Herceptin in solutions by using a small (2.2 kDa), inexpensive, highly stable human epidermal growth factor receptor (HER2) mimotope-derived synthetic peptide immobilized on the surface of a Au quartz electrode. We used the HER2 mimotope as a substitute for the HER2 receptor protein in piezoimmunosensor or quartz crystal microbalance (QCM) assays to detect Herceptin in human serum. We demonstrated that assay sensitivity was dependent upon the amino acids used to tether and link the peptide to the sensor surface and the buffers used to carry out the assays. The detection limit of the piezoimmunosensor assay was 0.038 nM with a linear operating range of 0.038-0.859 nM. Little nonspecific binding to other therapeutic antibodies (Avastin and Rituxan) was observed. Levels of Herceptin in serum samples obtained from treated patients, as ascertained using the synthetic peptide-based QCM assay, were typical for those treated with Herceptin. The findings of this study are significant in that low-cost synthetic peptides could be used in a QCM assay, in lieu of native or recombinant antigens or capture antibodies, to rapidly detect a therapeutic antibody in human serum. The results suggested that a synthetic peptide bearing a particular functional sequence could be applied for developing a new generation of affinity-based immunosensors to detect a broad range of clinical biomarkers.