摘要
为了延长IFNα2b在血浆中的半衰期,构建了编码HSA和hIFNα2b的融合基因并在毕赤酵母中获得高效表达,工程菌经5L发酵罐培养后获得的含融合蛋白的培养液经超滤浓缩、蓝色葡聚糖凝胶层析、疏水柱层析以及阴离子柱层析,融合蛋白的纯度达到95%以上。该融合蛋白能与干扰素抗体和人血清白蛋白抗体结合,并表现出与重组干扰素α2b相似的抗病毒活性。以猕猴为动物模型,分别从静脉和皮下单剂量给药,给药浓度为90μg/kg时,在336h后血浆中仍可检测到融合蛋白。其静脉注射的血浆半衰期为101h,皮下注射的半衰期为68.2h。皮下注射的生物利用度为67.9%。IFNα2b与HSA融合后,明显的延长了血浆半衰期,显现了其良好的临床应用前景。
To reduce the serum clearance of interferon α2b, a chimeric gene encoding an human serum albumin（HSA） - human interferon α2b（IFNα22b） fusion protein was overexpressed in Pichia pastoris. After fermentation in a 5L bioreactor, the fusion protein, capable of cross-reacting with anti-IFN α and anti-HSA antibody, was purified from the culture of the recombinant yeast by uhrafihration, blue Sepharose affinity, phenyl hydrophobic interaction and Q ion exchange chromatography. Its IFNα2b moiety exhibits antiviral activity similar to that of recombinant human IFNα2b. In Cynomolgus monkeys model, The fusion protein was detectable in plasma, even 336h after a single does of 90μg/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 α2b fused to human serum albumin suggest its promissing application in clinic medicine.
出处
《生物工程学报》
CAS
CSCD
北大核心
2006年第2期173-179,共7页
Chinese Journal of Biotechnology
