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Currently, a variety of display technologies have been developed in the life science field, such as phage display, ribosome display, and yeast display. Many studies have found that display technologies are powerful and universal methods when they are combined with large genetically encoded binder libraries, which results in the generation of high-performance binders against nearly any antigen interested. As a result, display technologies are widely applied to molecular biology, clinic and medicine. Ribosome display based on cell-free display systems has been established as a different type of technology for 23 years until now. Compared to other related methods, ribosome display possesses unique advantages and is successfully exploited to the selection for functional and specific binders in vitro, exhibiting potent development prospects. Here in this paper, we will review the theories and advantages of ribosome display, and will highlight how it is being used now to select and evolve functional proteins as well as applications in diagnostics and therapeutics.