The invention discloses a charge-induced sulphophile expanded adsorbent bed medium with hydrophobic nature and a preparation method thereof. In the composition of the medium, matrix is a composite microsphere with fibrin/inorganic weighting agent, petunidin is sulfonyl group and mercapto benzimidazole. The composite microsphere with fibrin/inorganic weighting agent is acquired by inverse suspension thermal regeneration and is taken as the matrix to mix divinyl sulfone, soda buffer solution and DMSO for activation, then swab-off active matrix, the mercapto benzimidazole and sodium hydroxide containing ammonium persulfate are mixed for coupling, thus acquiring the charge-induced sulphophile expanded adsorbent bed medium with the hydrophobic nature. The expanded bed adsorbing medium developed by the invention combines with three protein separation principles of hydrophobic charge induced chromatography, thiophilic chromatography and expanded bed adsorption, etc., which has the advantagesof large density of the petunidin, large protein adsorption capacity, strong specificity, mild conditions of adsorption and elution, etc., and shows the characteristic of the integration of technology and process, thus being used for high efficient separation and purification of proteins such as antibody, etc.