With growing global attention to food safety and sustainability, the development and application of novel protein sources—such as insect proteins and plant-based proteins—has become a research hotspot. These emerging protein sources offer diversified options for global protein supply; however, evaluating their allergenicity is a critical step in the development process. By combining in vitro and in vivo experiments with immunological responses from allergic individuals, the potential allergenic risks of novel proteins can be effectively assessed to ensure their safety for consumption.