Expanded bed adsorption (EBA), a promising and practical separation technique, has been widely studied in the past two decades. The development of adsorbents for EBA process is a challenging course, with the special design and preparation according to the target molecules and specific expanded bed systems. Many types of supporting matrices for expanded bed adsorbents have been developed, and their preparation methods are being consummated gradually. These matrices are activated and then coupled with ligands to form functionalized adsorbents, including ion-exchange adsorbents, affinity adsorbents, mixed mode adsorbents, hydrophobic charge induction chromatography adsorbents and others. In this review, the preparation of the matrices for EBA process is summarized, and the coupling of ligands to the matrices to prepare functionalized adsorbents is discussed as well.