The single molecule level determination with a transistor (SiMoT) platform has attracted considerable attention in the recognition of various ultralow abundance biomolecules, while complicated labeling and testing processes limit its further applications. Recently, organic field-effect transistor (OFET)-based biosensors are good candidates for constructing an advanced label-free SiMoT platform due to their facile fabrication process, rapid response time, and low sample volume with a wide range of detection. However, the sensitivity of most OFET-based biosensors is in the order of nM and pM, which cannot meet the detection requirements of ultralow abundance protein. Herein, a label-free SiMoT platform is demonstrated by integrating pillar[n]arene as a signal amplifier, and the detection limit can reach 4.75 aM. Besides, by simultaneous determination of α-fetoprotein, carcinoembryonic antigen, and prostate antigen, the proposed multiplexed OFET-based SiMoT platform provides a key step in reliable early cancer diagnosis.