Expression and characterization of recombinant human serum albumin fusion protein with C-peptide
| Autor: | Yang, HL, Wei, YY, Zhou, LP, Kong, JJ |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
C-peptide
human serum albumin recombinant fusion protein Pichia pastoris bioactivity biological half-time C-peptide Human serum albumin Applied Microbiology and Biotechnology Molecular biology Fusion protein law.invention chemistry.chemical_compound chemistry law Genetics medicine Recombinant DNA Agronomy and Crop Science Molecular Biology Biotechnology medicine.drug |
| Zdroj: | African Journal of Biotechnology; Vol 10, No 71 (2011); 16058-16063 |
| ISSN: | 1684-5315 |
| DOI: | 10.5897/ajb11.643 |
| Popis: | C-peptide (CP), connecting the A and B chains in proinsulin, has been considered to possess physiological effects in diabetes. In order to prolong the half-life of CP in vivo, a long acting CP analog [human serum albumin (HSA-CP)] was obtained by direct gene fusion of a single-chain CP to HSA and expressed in host Pichia pastoris GS115. After 72 h of growth on methanol, the recombinant HSA-CP concentration reached a level of 145 mg/L, representing 70% of total proteins in culture supernatant. The recombinant fusion protein was purified by ultra filtration, Q-sepharose fast flow column and Superdex 75 size-exclusion column. It was specifically recognized by the anti-human HSA antibody and CP antibody in Western blotting assay. In vitro, the recombinant HSA-CP can stimulate HEK293 cell proliferation. In Zucker diabetic fatty (ZDF) rats, 12 weeks recombinant HSA-CP treatment could prevent the accumulation of glomerular extracellular matrix, which leads to mesangial expansion and glomerular hypertrophy. The terminal biological half-time of the protein was 3.38±1.87 h after single administration of 500 nmol/kg of HSA-CP in Wister rats. The pharmacokinetic analysis of the protein indicate its promising application in clinical medicine.Key words: C-peptide, human serum albumin, recombinant fusion protein, Pichia pastoris, bioactivity,biological half-time. |
| Databáze: | OpenAIRE |
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