Lectins are a glycoprotein that plays a significant role in plant and fungi resistance mechanism against insect and microorganism attacks. This paper describes for the first time the extraction and purification of a lectin protein found in the tubers of Truffle (Terfezia sp.) by a series of steps. Initially, precipitated by ammonium sulfate and accompanied by chromatographic techniques include ion-exchange chromatography and gel permeation chromatography. Lectin proteins were showed purification fold about 20.851 in the final step of the purification and specific activity about 377.581 unit/mg protein. Also, the hemagglutination activity assay of the Truffle lectin was showed activity to agglutinate for each step with the blood types A and O more than B group at room temp, under moderate pH 7.0 and also show high stability temperature extend from 0°to 40°C based on the hemagglutination activity in repeated experiments under various temperatures. The carbohydrates specificity assay results showed that the lectins have complex specificities for many sugars, but not have specific for another group in this test.