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Immunotherapy using CAR-T cells is a new paradigm technology for cancer treatment. To avoid severe side effects and tumor escape variants observed for conventional CAR-T cells approach, adaptor CAR technologies are under development, where intermediate target modules redirect immune cells against cancer cells. In this work, silicon nanowire field effect transistors are used to assist in the development of target modules for an optimized CAR-T cell operation. Focusing on a library of seven variants of E5B9 peptide that is used as CAR peptide epitope, we performed multiplexed binding tests in serum using nanosensor chips. Peptides have been immobilized onto the sensor to compare the signals of transistor upon titration with anti-E5B9 antibodies. Correlation analysis of binding affinities and sensitivities enabled a selection of best candidates for the interaction between CAR and target modules. Finally, cytotoxic functionality of CAR-T cells in combination with the selected target modules were successfully proven. Our results open the perspective for the nanobiosensorics to go beyond the early diagnostics in the field of clinical cancer research, and paves the way towards personalization and efficient monitoring of the immunotherapeutic treatment, where the quantitative analysis with the standard techniques is not an option.