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Alpha-fetoprotein (AFP) is an important disease biomarker, related to cancers such as hepatocarcinomas and gastric cancer. However, traditional methods are time-consuming, relied on bulky instruments and trained professionals, cannot satisfy the demand for low cost and point-of-care testing (POCT). In this study, a power-free POCT device is developed for rapid and low-cost detection of AFP via one-sampling. Based on the principle of sandwich immunofluorescence, the chip is capable of automatically accomplishing on-chip mixing, labeling and capturing procedures, which only requires the operator add 40 μl sample into the chip one time. The proposed device is capable of sensitively detecting human AFP in FBS with a dynamic ranging from 10-1 000 ng/mL and LOD (1.88 ng/ml) within a short time of 3 min.Predictably, our method holds the great potential to be applied in POC diagnostics of proteins, especially for some regions with resource-limited.