Extending the Serum Half-Life of G-CSF via Fusion with the Domain III of Human Serum Albumin

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...

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Bibliographic Details
Published in:BioMed research international 2013-01, Vol.2013 (2013), p.1-8
Main Authors: Gao, Xiangdong, Cai, Di, Chen, Xiaofei, Tian, Hong, Zhang, Yu, Zhao, Shuqiang, Yao, Wenbing
Format: Article
Language:English
Subjects:
Animals
Antibodies
Biological activity
Cloning
Enzymes
Gene Expression Regulation
Genetic engineering
Granulocyte Colony-Stimulating Factor - blood
Granulocyte Colony-Stimulating Factor - chemistry
Granulocyte Colony-Stimulating Factor - genetics
Half-Life
Humans
Laboratory animals
Leukocyte Count
Medical research
Mice
Microbiology
Molecular weight
Pichia
Pichia pastoris
Protein Structure, Tertiary - genetics
Proteins
Recombinant Fusion Proteins - chemistry
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - pharmacokinetics
Serum Albumin - chemistry
Serum Albumin - genetics
Studies
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Summary: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 pPICZαA 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.
ISSN:2314-6133
2314-6141
DOI:10.1155/2013/107238