[Expression in Pichia pastoris and properties of human serum albumin-interferon alpha2b chimera].
To reduce the serum clearance of interferon alpha2b, a chimeric gene encoding an human serum albumin(HSA)--human interferon alpha2b(IFNalpha2b) fusion protein was overexpressed in Pichia pastoris. After fermentation in a 5L bioreactor, the fusion protein, capable of cross-reacting with anti-IFN alpha and anti-HSA antibody, was purified from the culture of the recombinant yeast by ultrafiltration, blue Sepharose affinity, phenyl hydrophobic interaction and Q ion exchange chromatography. Its IFNa2b moiety exhibits antiviral activity similar to that of recombinant human IFNa2b. In Cynomolgus monkeys model, The fusion protein was detectable in plasma, even 336h after a single does of 90 microg/kg injection intravenously or subcutaneously. The elimination phase half-life of the fusion protein was 101h after intravenous injection and 68.2h after subcutaneous injection. Its Subcutaneous bioavailability was 67.9%. The enhanced pharmacokinetics of interferon a2b fused to human serum albumin suggest its promissing application in clinic medicine.
Chang
Shao-Hong
SH
Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China.
Gong
Xin
X
Yang
Zhi-Yu
ZY
Wang
Tong-Ying
TY
Ma
Guo-Chang
GC
Ma
Qing-Jun
QJ
Wu
Jun
J
D000818:Animals; D019149:Bioreactors / Q000382:microbiology; D005285:Fermentation; D006801:Humans; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000096:biosynthesis / Q000235:genetics*; D008252:Macaca fascicularis; D010843:Pichia / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000493:pharmacokinetics*; D011994:Recombinant Proteins; D012709:Serum Albumin / Q000096:biosynthesis / Q000235:genetics*
D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D012709:Serum Albumin