The raw materials of Poly(ethylene terephthalate) (PET) are derived from petroleum‐based resources, which are no sustainable. Therefore, previous researchers introduced biomass‐derived 2,5‐tetrahydrofurfuryl dimethanol (THFDM) into PET. However, its heat resistance has decreased compared to PET. In this paper, a novel bio‐based copolyester, poly(ethylene glycol‐co‐2,5‐tetrahydrofuran dimethanol‐co‐isosorbide terephthalate) (PEIFT), is prepared by introducing biomass‐derived isosorbide (ISB) and THFDM into the PET chains through melting copolymerization process. With the introduction of ISB content, copolyesters' hydrophilicity and rigidity improve. Compared to PET, glass transition temperature (Tg) increases by over 5 °C. In addition, the toughness and spinning performance of PEIFT have also been improved as a result of the addition of THFDM components. The hydrophobicity of PEIFTs electrospinning is greatly improved, with a contact angle exceeding 135 °. Finally, due to the good hydrophobicity of PEIFTs nanofibers, they have potential application value in the manufacture of hydrophobic nanofiber and filter films. Given its biomass source and excellent performance, they make it easier to replace materials derived from petroleum.This article is protected by copyright. All rights reserved