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Background: Enzymes are natural protein catalysts that accelerate biochemical reactions in food, pharmaceutical, and industrial processes, but their free (soluble) form is often unstable and difficult to recover and reuse. Objective: This review provides an overview of enzyme immobilization technology, covering the support materials and techniques used to immobilize enzymes, and their applications in biosensing, food safety, and medical diagnostics. Methods: Peer-reviewed literature on enzyme immobilization was reviewed and synthesized, focusing on organic and inorganic support systems, immobilization techniques (adsorption, covalent binding, entrapment, encapsulation, and cross-linking), and reported applications of immobilized enzymes and enzyme-based biosensors. Results: Support selection and immobilization technique together determine enzyme stability, reusability, and catalytic performance. Enzyme-based biosensors built on these principles have been reported for food-quality monitoring, contaminant and pesticide detection, glucose monitoring in diabetes, and cardiac biomarker detection in cardiovascular disease. Conclusion: Enzyme immobilization is a versatile and cost-effective strategy that improves enzyme stability and reusability across pharmaceutical, biomedical, food, and industrial applications, with continued advances in support materials and nanotechnology expected to expand its future applications.