烟草细胞中高效表达提高血清半衰期的药用蛋白  

High-yield Expression of Therapeutic Proteins with Extended Serum Half-life in Tobacco Cells

作　　者：Jianfeng Xu Marcia J. Kieliszewski 

机构地区：[1]Arkansans Biosciences Institute, Arkansans State University, State College, AR 72467, USA [2]Department of Chemistry and Biochemistry, Ohio University, Athens, OH 45701, USA

出　　处：《生物工程学报》2008年第12期2130-2130,共1页Chinese Journal of Biotechnology

摘　　要：小分子治疗蛋白，如干扰素α2b（IFNα2）和人生长激素（hGH），由于其对血清蛋白酶的易感性及分子小，可被肾迅速清除，因而一般在血清中的半衰期都很短。化学衍生处理虽可以克服这些问题，但却要以降低蛋白的生物活性为代价。本研究建立了一种新方法，使干扰素仅2b（IFNα2）和人类生长激素（hGH）在烟草细胞中高效表达为阿拉伯半乳糖聚糖糖蛋白嵌合体（AGP）。这样不仅提高了药用蛋白的产量，而且提高了其血清半表期。这种嵌合糖蛋白的分泌量比没有糖基化时高500～1800倍。重要的是，与未糖基化的干扰素和人生长激素相比，这种嵌合糖蛋白的体内血清半衰期提高13倍，生物活性仍保持与未糖基化时相似。这种嵌合糖蛋白注射到小鼠体内未引起免疫性反应。Small therapeutic proteins such as interferon α2b （IFNα2） and human growth hormone （hGH） generally possess short serum half-lives due to their susceptibility to serum proteases and small size, hence rapid renal clearance. Chemical derivatization overcomes these problems but at the expense of decreased bioactivity. We developed a new method that yields biologically IFNα2 and hGH in high yields and with increased serum half-lives when expressed as arabinogalactan-protein （AGP） chimeras in cultured tobacco cells. The chimeric glycoproteins typically gave 500-1800 fold greater secreted yields than their non-glycosylated controls. Importantly, the chimeric glycoproteins showed an increased in vivo serum half-life of up to 13 fold while their biological activities remain similar to native IFNα2 and hGH. The AGP domain was not immunogenic when injected into mice.

关 键 词：植物细胞培养 重组蛋白 糖基化羟脯胺酸 半衰期 

分 类 号：Q78[生物学—分子生物学] S828.2[农业科学—畜牧学]