Proteins are the executors of various kinds of metabolism and regulation in cells and they are also the most important target molecules of pathogenic factors and drugs. The study of protein expression is necessary to understand life, disease processes and drug effects. At present, routine protein detection methods in clinical practice require the support of large equipment. However, with the development of medical technologies and especially under the special background of the coronavirus (COVID-19) pandemic, point-of-care testing (POCT, also known as on-site testing and bedside testing) has become the current development trend. POCT can improve the ways of interaction between patients and doctors and create a positive approach to medical treatment. In addition to diagnosing and treating diseases, POCT has advantages in both on-site and remote detection for personnel engaged in emergency work. Therefore, it is very important to develop accurate, sensitive, simple and fast protein POCT. To develop miniature devices for POCT, many new approaches have been attempted in recent years, including microfluidics, electrochemical biosensors, smart phones, artificial intelligence, and wearable devices. Microfluidics deals with small quantities of samples and is a universal platform for integrating a variety of technologies, such as immunochemistry, electrochemistry, and mass spectrometry. With microfluidic technology, the sample size, reaction time, and detection limit could be easily improved to satisfactory levels. The use of electrochemistry in detecting proteins has opened a new field in POCT. Since the core of electrochemistry lies in the nanocatalyst development, the rapidly growing research on nanomaterials also has facilitated and expanded the areas of POCT applications. Furthermore, the current frontier technologies employing 5G, artificial intelligence, and wearable devices have not only generated new possibilities but also greatly inspired scientists to create novel POCT devices. In conclusion, with the increasing demand of people and the continuous progress of science and technology, the development direction of future protein detection methods will be focused on portability, intellectualization, rapidity, integration of diagnosis and treatment by multidisciplinary approaches for better application in clinical practice and applicability for bedside and home testing.