Natural polysaccharides are macromolecular substances with a wide range of biological activities. The structural modification of polysaccharides by chemical means can enhance their biological activity. This paper reviews the latest research reports on the chemical modification of natural polysaccharides. At present, the modification methods of polysaccharides mainly include sulfation, phosphorylation, carboxymethylation, socialization, methylation and acetylation. The chemical and physical structures of the modified polysaccharides were detected via ultraviolet spectroscopy, FT-IR, high-performance liquid chromatography, ultraviolet spectroscopy, gas chromatography–mass spectrometry, nuclear magnetic resonance and scanning electron microscopy. Modern pharmacological studies have shown that the modified polysaccharide has various biological activities, such as antioxidant, antitumor, immune regulation, antiviral, antibacterial and anticoagulant functions in vitro. This review provides fresh ideas for the research and application of polysaccharide structure modification.