Expression, purification and characterization of recombinant human interleukin-2-serum albumin (rhIL-2-HSA) fusion protein in Pichia pastoris

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• 作者：Jianyong Lei^a ; ¹ ; ^{lyreau@126.com} ; Bo Guan^a ; ¹ ; ^{gbb3519@yahoo.com.cn} ; Bo Li^a ; Zuoying Duan^a ; Yun Chen^b ; Huazhong Li^a ; ^{hzhli@jiangnan.edu.cn} ; Jian Jin^a ; ^b ; ^{jinjian31@126.com}
• 关键词：Human interleukin-2 ; Human serum albumin ; Pichia pastoris ; Secretory expression ; Fusion protein
• 刊名：Protein Expression and Purification
• 出版年：2012
• 期刊代码：184_10465928
• 类别：bio
• 出版时间：July, 2012
• 卷：84
• 期：1
• 页码：154-160
• 文件大小：587 K

摘要

Interleukin-2 (IL-2) plays important roles in variety of immune functions. Recombinant IL-2 has become an important therapeutic protein for therapy of melanoma and renal cell carcinoma. Previously, it was proved that the therapeutic efficacy of rIL-2 expressed in Saccharomyces cerevisiae was improved by prolonging its in vivo half-life through genetic fusion with albumin. In this study, a fusion protein composed of hIL-2 genetically fused to HSA was expressed as a secretory protein under AOX1 (alcohol oxidase 1) promoter in Pichia pastoris. An effective strategy was established to express rhIL-2-HSA fusion protein in 5 L scale and the optimal purification procedure was investigated. The purity of rhIL-2-HSA in final product was about 95%. The purified rhIL-2-HSA fusion protein could be recognized by both anti-hIL-2 and anti-human serum albumin monoclonal antibody. Bioactivity analysis showed that the purified rhIL-2-HSA fusion protein displayed high level activity on proliferation in IL-2 dependent manner in CTLL2-cells. rhIL-2-HSA fusion protein also showed a extended half-life in plasma compared with IL-2 when tested in a BALB/c mouse model. This study provides an alternative strategy for large-scale production of bioactive rhIL-2-HSA fusion protein using P. pastoris as an expression host.