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当前位置: 首页 > 文章 > 昆虫细胞内N-糖基化途径及人源化糖蛋白表达 昆虫学报 2011,54 (6) 112-118
Position: Home > Articles > Protein N-glycosylation pathway and expression of humanized glycoproteins in insect cells Acta Entomologica Sinica 2011,54 (6) 112-118

昆虫细胞内N-糖基化途径及人源化糖蛋白表达

作  者：
胡嘉彪;张芃;牛艳山;Bhaskar ROY;李兴华;杨华军;周芳;缪云根
单  位：
浙江大学动物科学学院
关键词：
昆虫细胞;哺乳动物细胞;糖蛋白;蛋白糖酰化途径;糖苷;人源化糖蛋白
摘  要：
虽然昆虫杆状病毒表达系统在蛋白表达领域得到了广泛的应用,但由于不能表达复杂的末端唾液酸化的N-糖链,使得该系统在生物制药行业的应用受到了很大的限制。通过比较哺乳动物细胞和昆虫细胞内糖基化途径可知,其起始步骤一致,之后再发生分化,主要表现为3方面,即昆虫细胞内缺乏哺乳动物细胞所具备的N-乙酰葡萄糖氨转移酶II、半乳糖基转移酶/N-乙酰氨基半乳糖转移酶、α-2,3-唾液酸转移酶和α-2,6-唾液酸转移酶等延长N-糖链的糖基转移酶;另外,昆虫细胞内具有能够特异性地将蛋白质末端的N-乙酰氨基葡萄糖残基从GlcNAcMan3GlcNAc(±α3/6-Fuc)GlcNAc上切除的N-乙酰氨基葡萄糖苷酶及核心α-1,3-岩藻糖基转移酶。本文从上述异同出发,综述了克服昆虫细胞内不能表达人源化糖蛋白这一缺陷所进行的N-糖基化途径的改造研究——主要集中在昆虫细胞内GlcNAcase的抑制和昆虫细胞内GnT2,GalT/GalNAcT,ST3及ST6等基因的导入等方面,结果表明经改造的昆虫细胞可表达人源化糖蛋白,这将极大地拓宽昆虫杆状病毒表达系统的应用领域。本文还探讨了选择特殊细胞系及特殊培养条件以在昆虫细胞内表达唾液酸化蛋白的可行性。
译  名：
Protein N-glycosylation pathway and expression of humanized glycoproteins in insect cells
作  者：
HU Jia-Biao,ZHANG Peng,NIU Yan-Shan,Bhaskar ROY,LI Xing-Hua,YANG Hua-Jun,ZHOU Fang,MIAO Yun-Gen (College of Animal Sciences,Zhejiang University,Hangzhou 310029,China)
关键词：
Insect cells;mammalian cells;glycoprotein;protein N-glycosylation pathway;glycan;humanized glycoprotein
摘  要：
Insect-baculovirus expression system has been widely used to produce recombinant proteins.However,due to its deficiency in the post-translation modification,complex terminal sialylated N-linked glycans can not be obtained,thereby largely limiting the application of this system in biopharmaceutical industry.By comparing the glycosylation pathways in mammalian and insect cells,it is known that the initial steps are the same and then diverge.The differences mainly include that insect cells lack the mammalian glycosyltransferases like N-acetylglucosaminyltransferase II,galaetosyltransferase/N-acetylgalactosyltransferase,α-2,3-sialyltransferase and α-2,6-sialyltransferase.On the other hand,insect cells possess specific α-1,3-fucosyltransferase and N-acetylglucosaminidase which can specifically remove the terminal GlcNAc from GlcNAcMan3GlcNAc(±α3/6-Fuc)GlcNAc.Based on the comparison,this article summarized the research efforts to produce humanized proteins in insect cells through the inhibition of GlcNAcase and knock-in of the mammalian glycosyltransferases.The results showed that engineered insect cells could produce humanized glycoproteins,which would greatly expand the application of insect-baculovirus expression system.In addition,the feasibility of production of humanized proteins by selection of novel insect cell lines and/or culture condition was discussed.

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