The invention provides a colloidal crystal adsorption chromatographic separation method for lysozyme, which comprises the following steps of: 1. diluting mixed liquid containing lysozyme with a buffer solution with pH value between 6.0-10.5 to obtain a loading solution, absorbing by using a colloidal crystal bed column at the flow velocity of 0.1-25cm/min; 2. washing off impurities in colloidal crystals, which are not absorbed, with deioned water or a buffer solution with pH value between 6.0-10.5; and 3. eluting with an eluent which is a buffer solution containing 0.02-3M of alkali metal salt and having pH value between 6.0-10.5, collecting an elution peak containing lysozyme to obtain the lysozyme. The method has the advantages of simple process and large processing capacity, and the purity of the lysozyme obtained by the method is high, the used colloidal crystal medium can be regenerated conveniently and can be reused, and thus, the method is a low-cost lysozyme separation and purification method.