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首页> 外文期刊>Biotechnology Progress >Improved Secretion of Molecular Chaperone-Assisted Human IgG in Silkworm,and No Alterations in Their N-Linked Glycan Structures
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Improved Secretion of Molecular Chaperone-Assisted Human IgG in Silkworm,and No Alterations in Their N-Linked Glycan Structures

机译：分子伴侣中人陪伴分子IgG的分泌得到改善，并且它们的N-连接聚糖结构无变化

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摘要

Human 29IJ6 IgG was expressed in silkworm using a Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid system. The mean amounts of 296IJ6 IgG produced in larval hemolymph and whole pupae were 30.1 μg/larva and 78.0 μg/pupa, respectively. The use of molecular chaperones including calreticulin (CRT), calnexin (CNX), and immunoglobulin heavy chain binding protein (BiP, GRP78) improved the production of 296IJ6 IgG secretion in the larvae fivefold. The total yield of recombinant 29IJ6 IgG was 239 μg/mL when coexpressed with CRT. However, the overexpression of molecular chaperones had negative effects on secretion. The N-linked glycans of secreted 296IJ6 IgG in silkworm hemolymph were dominated by paucimannose structures. Small amounts of GlcNAc residues linked to the Manα1,3 branch were detected. When molecular chaperones were coexpressed, the compositions of N-linked glycans in the IgG1 produced were unchanged compared with those produced without them. This suggests that N-glycosylation is controlled by a regulatory function in the Golgi apparatus even though the post-translational modification of 296IJ6 IgG was assisted by the coexpression of molecular chaperones. Therefore, if the glycosylation pathways that coexpress N-acetylglucosaminyltransferase, galactosyltransferase, and sialyltransferase could be improved, silkworm larvae might prove a useful system for producing human antibodies.
机译：使用家蚕核多角体病毒（BmNPV）杆粒系统在蚕中表达人29IJ6 IgG。幼虫血淋巴和整个p中产生的296IJ6 IgG的平均量分别为30.1μg/幼虫和78.0μg/ pupa。使用包括钙网蛋白（CRT），钙连接蛋白（CNX）和免疫球蛋白重链结合蛋白（BiP，GRP78）的分子伴侣可以将幼虫中296IJ6 IgG分泌的产生提高五倍。与CRT共表达时，重组29IJ6 IgG的总产量为239μg/ mL。然而，分子伴侣的过表达对分泌有负面影响。家蚕血淋巴中分泌的296IJ6 IgG的N-连接聚糖以paucimannose结构为主。检测到少量与Manα1,3分支连接的GlcNAc残基。当分子伴侣被共表达时，所产生的IgG1中的N-连接聚糖的组成与没有它们时所产生的那些相同。这表明即使296IJ6 IgG的翻译后修饰通过分子伴侣蛋白的共表达来辅助，高尔基体中的N-糖基化也受调节功能的控制。因此，如果可以改善共表达N-乙酰氨基葡糖基转移酶，半乳糖基转移酶和唾液酸转移酶的糖基化途径，则蚕幼虫可能证明是生产人抗体的有用系统。

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