title: Extending the serum half-life of G-CSF via fusion with the domain III of human serum albumin.
abstract: Protein fusion technology is one of the most commonly used methods to extend the half-life of therapeutic proteins. In this study, in order to prolong the half-life of Granulocyte colony stimulating factor (G-CSF), the domain III of human serum albumin (3DHSA) was genetically fused to the N-terminal of G-CSF. The 3DHSA-G-CSF fusion gene was cloned into pPICZ α A along with the open reading frame of the α -factor signal under the control of the AOX1 promoter. The recombinant expression vector was transformed into Pichia pastoris GS115, and the recombinant strains were screened by SDS-PAGE. As expected, the 3DHSA-G-CSF showed high binding affinity with HSA antibody and G-CSF antibody, and the natural N-terminal of 3DHSA was detected by N-terminal sequencing. The bioactivity and pharmacokinetic studies of 3DHSA-G-CSF were respectively determined using neutropenia model mice and human G-CSF ELISA kit. The results demonstrated that 3DHSA-G-CSF has the ability to increase the peripheral white blood cell (WBC) counts of neutropenia model mice, and the half-life of 3DHSA-G-CSF is longer than that of native G-CSF. In conclusion, 3DHSA can be used to extend the half-life of G-CSF.
authors.affiliation: State Key Laboratory of Natural Medicines, School of Life Science and Technology, China Pharmaceutical University, Tongjiaxiang 24, Nanjing 210009, China.
authors.forename: Shuqiang
authors.initials: S
authors.lastname: Zhao
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.forename: Hong
authors.initials: H
authors.lastname: Tian
authors.forename: Xiaofei
authors.initials: X
authors.lastname: Chen
authors.forename: Di
authors.initials: D
authors.lastname: Cai
authors.forename: Wenbing
authors.initials: W
authors.lastname: Yao
authors.forename: Xiangdong
authors.initials: X
authors.lastname: Gao
mesh_terms: D000818:Animals; D000906:Antibodies; D005786:Gene Expression Regulation; D016179:Granulocyte Colony-Stimulating Factor / Q000097:blood / Q000737:chemistry / Q000235:genetics*; D006207:Half-Life; D006801:Humans; D007958:Leukocyte Count; D051379:Mice; D010843:Pichia; D017434:Protein Structure, Tertiary / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics* / Q000493:pharmacokinetics; D012709:Serum Albumin / Q000737:chemistry / Q000235:genetics*
chemical_list: D000906:Antibodies; D011993:Recombinant Fusion Proteins; D012709:Serum Albumin; D016179:Granulocyte Colony-Stimulating Factor