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This invention relates to a method for improving protein crystallization efficiency using hydrogel regulation. The method includes the following steps: mixing monomers to obtain a prepolymer solution, adding water and a photoinitiator, stirring to obtain a hydrogel precursor solution; stabilizing the obtained hydrogel precursor solution, then irradiating it with ultraviolet light to obtain a rough hydrogel; immersing the rough hydrogel in water and then drying it to obtain a dried hydrogel; preparing a buffer solution and using the prepared buffer solution to prepare a precipitant; immersing the dried hydrogel obtained in the previous step in the prepared precipitant to swell the dried hydrogel, washing and drying it to obtain a target hydrogel; dissolving the protein in the prepared buffer solution to obtain a protein solution; and dropping the obtained protein solution onto the surface of the obtained target hydrogel to crystallize it to obtain protein crystals. Compared with existing technologies, this invention can effectively solve the problem of low efficiency in protein crystallization while ensuring the quality of protein crystals.