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Engineering of CHO Cells for the Production of Recombinant Glycoprotein Vaccines with Xylosylated N -glycans

机译:具有木糖基化N-聚糖的重组糖蛋白疫苗生产的CHO细胞工程化

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Xylose is a general component of O -glycans in mammals. Core-xylosylation of N -glycans is only found in plants and helminth. Consequently, xylosylated N -glycans cause immunological response in humans. We have used the F-protein of the human respiratory syncytial virus (RSV), one of the main causes of respiratory tract infection in infants and elderly, as a model protein for vaccination. The RSV-F protein was expressed in CHO-DG44 cells, which were further modified by co-expression of β1,2-xylosyltransferase from Nicotiana tabacum . Xylosylation of RSV-F N -glycans was shown by monosaccharide analysis and MALDI-TOF mass spectrometry. In immunogenic studies with a human artificial lymph node model, the engineered RSV-F protein revealed improved vaccination efficacy.
机译:木糖是哺乳动物中O-聚糖的一般成分。 N-聚糖的核心木糖基化仅在植物和蠕虫中发现。因此,木糖基化的N-聚糖在人中引起免疫应答。我们已经使用了人类呼吸道合胞病毒(RSV)的F蛋白作为疫苗模型蛋白质,RSV是婴幼儿呼吸道感染的主要原因。 RSV-F蛋白在CHO-DG44细胞中表达,并通过烟草的β1,2-木糖基转移酶的共表达进一步修饰。通过单糖分析和MALDI-TOF质谱显示了RSV-F N-聚糖的木糖基化。在使用人类人工淋巴结模型的免疫原性研究中,经工程改造的RSV-F蛋白显示出更高的疫苗接种效力。

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