title: Functional expression of human serum albumin-tandem thrombopoietin mimetic peptide fusion protein as a novel thrombopoietin analog in Pichia pastoris.
abstract: OBJECTIVE
  To develop a novel thrombopoietin (TPO) analog by fusing the tandem TPO mimetic peptide (TMP-TMP) to human serum albumin (HSA) and performing functional expression of recombinant fusion protein HSA-TMP-TMP.
  
  
  RESULTS
  After optimizing the fusion orientation in shake-flask culture, HSA-TMP-TMP was expressed at 0.4 g/l in Pichia pastoris grown in a 20 l bioreactor, during which pH was controlled at 5 by addition of NH4OH and citric acid. The fusion protein significantly activated signal transducer and activator of transcription-mediated transcription in TPO receptor-dependent manner, which was demonstrated by a luciferase reporter assay. Following subcutaneous administration, HSA-TMP-TMP effectively stimulated the platelet production in healthy mice in a dose-dependent manner.
  
  
  CONCLUSIONS
  Successful expression of HSA-TMP-TMP fusion protein in P. pastoris was achieved and the recombinant HSA-TMP-TMP is a promising TPO analog.
authors: Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, Institute of Microbiology, School of Life Sciences, Lanzhou University, Tianshui South Road No. 222, Lanzhou, 730000, People's Republic of China.
authors: Meizhu
authors: M
authors: Wang
authors: Gansu High Throughput Screening and Creation Center for Health Products, School of Pharmacy, Lanzhou University, Lanzhou, People's Republic of China.
authors: Dejuan
authors: D
authors: Zhi
authors: Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, Institute of Microbiology, School of Life Sciences, Lanzhou University, Tianshui South Road No. 222, Lanzhou, 730000, People's Republic of China.
authors: Jun
authors: J
authors: Xian
authors: Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, Institute of Microbiology, School of Life Sciences, Lanzhou University, Tianshui South Road No. 222, Lanzhou, 730000, People's Republic of China.
authors: Yi
authors: Y
authors: Ru
authors: Gansu High Throughput Screening and Creation Center for Health Products, School of Pharmacy, Lanzhou University, Lanzhou, People's Republic of China.
authors: Suzhen
authors: S
authors: Wei
authors: Gansu High Throughput Screening and Creation Center for Health Products, School of Pharmacy, Lanzhou University, Lanzhou, People's Republic of China.
authors: Na
authors: N
authors: Wang
authors: Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, Institute of Microbiology, School of Life Sciences, Lanzhou University, Tianshui South Road No. 222, Lanzhou, 730000, People's Republic of China.
authors: Yiyao
authors: Y
authors: Liu
authors: Gansu High Throughput Screening and Creation Center for Health Products, School of Pharmacy, Lanzhou University, Lanzhou, People's Republic of China.
authors: Haiqing
authors: H
authors: Wang
authors: Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, Institute of Microbiology, School of Life Sciences, Lanzhou University, Tianshui South Road No. 222, Lanzhou, 730000, People's Republic of China.
authors: Yuejuan
authors: Y
authors: Pei
authors: Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, Institute of Microbiology, School of Life Sciences, Lanzhou University, Tianshui South Road No. 222, Lanzhou, 730000, People's Republic of China.
authors: Meijuan
authors: M
authors: Song
authors: Gansu High Throughput Screening and Creation Center for Health Products, School of Pharmacy, Lanzhou University, Lanzhou, People's Republic of China.
authors: Yang
authors: Y
authors: Li
authors: Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, Institute of Microbiology, School of Life Sciences, Lanzhou University, Tianshui South Road No. 222, Lanzhou, 730000, People's Republic of China. hyli213@yahoo.com.|Gansu High Throughput Screening and Creation Center for Health Products, School of Pharmacy, Lanzhou University, Lanzhou, People's Republic of China. hyli213@yahoo.com.
authors: Hongyu
authors: H
authors: Li
mesh_terms: D000818:Animals; D019149:Bioreactors; D015870:Gene Expression; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007279:Injections, Subcutaneous; D051379:Mice; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D010843:Pichia / Q000235:genetics / Q000378:metabolism; D053628:Receptors, Thrombopoietin / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D012709:Serum Albumin / Q000235:genetics / Q000378:metabolism*; D016896:Treatment Outcome
chemical_list: D010455:Peptides; D053628:Receptors, Thrombopoietin; D011993:Recombinant Fusion Proteins; D012709:Serum Albumin; C515728:thrombopoietin mimetic peptide
keywords: Fusion orientation; Human serum albumin-tandem thrombopoietin mimetic peptide fusion protein; Luciferase reporter assay; Pichia pastoris; Platelet count; Thrombocytopenia