Vanillin is an important flavoring agent that is mainly obtained from the seedpods of the tropical orchid, Vanilla planifolia. Molecularly imprinted polymer microspheres (MIPs) were synthesized by precipitation polymerization, suspension polymerization, and bulk polymerization using vanillin as the template molecule, methacrylic acid (MAA) as the functional monomer, ethylene glycol dimethacrylate (EGDMA) as the cross-linker, and dimethyl sulfoxide as the solvent. The interaction between vanillin and MAA of vanillin in MIPs and the best molar ratios were studied using UV spectroscopy and Fourier transform infrared spectroscopy (FT-IR). In addition, their microstructure was observed by transmission electron microscopy (TEM). Equilibrium adsorption and isothermal adsorption experiments were carried out to investigate the vanillin-binding capacities of the polymers. The results indicated that MAA and vanillin interacted via hydrogen bonding. Furthermore, the MIPs synthesized by precipitation polymerization were regularly shaped and had relatively high specificity for the recognition of vanillin.