Pretreatment and fractionation are amongst the key steps for the conversion of lignocelluloses to sustainable biofuels, biomaterials or biochemicals, as pretreatment/fractionation can break the natural recalcitrance of lignocelluloses, improving the conversion efficiency of downstream processes. This paper reviews the recent progress on the pretreatment and fractionation of lignocelluloses for biorefinery at the Chinese Academy of Sciences-Qingdao Institute of Bioenergy and Bioprocess Technology (QIBEBT). The main technologies developed at the QIBEBT in recent years include alkaline twin-screw extrusion pretreatment, modified alkali pretreatment, hydrogen peroxide-assisted sodium carbonate pretreatment, fractionation with formic acid, as well as the two-step fractionation by hot water treatment coupling ammonium sulfite treatment. With the development of these technologies, a pilot scale platform for the pretreatment and saccharification of biomass has been established in the pilot plant of QIBEBT.