The invention relates to the technical field of 
in vitro diagnostic reagents, and particularly relates to a lyophilized 
heparanase preparation, a 
heparanase cup as well as a preparation method and application of the lyophilized 
heparanase preparation and the heparanase cup. A redissolved 
enzyme activity yield of the lyophilized heparanase preparation provided by the invention is at least 75 percent. The heparanase cup provided by the invention comprises a cup body and the lyophilized heparanase preparation, wherein an accommodating space is defined in the cup body; the lyophilized heparanase preparation is arranged in the accommodating space. The invention also provides a method for preparing the lyophilized heparanase preparation. The method comprises the following steps: sequentially performing pre-freezing treatment, sublimation treatment and 
drying treatment on aqueous solutions prepared from 
heparinase, a protective agent, buffer salt and a 
preservative in a predetermined proportion so as to obtain the lyophilized heparanase preparation. The lyophilized heparanase preparation and the heparanase cup are used in a detection process of blood samples, and the detection sensitivityis high; moreover, the activity and the stability of the 
heparinase in a lyophilization process can be obviously improved.