Production of recombinant IgA1 with defined mucin-type O-glycans in Nicotiana tabacum BY-2 cells.

IF 4.1 2区 生物学 Q1 PLANT SCIENCES
Frontiers in Plant Science Pub Date : 2025-09-24 eCollection Date: 2025-01-01 DOI:10.3389/fpls.2025.1675517
Nicolas Bailly, Clemens Grünwald-Gruber, Marie Peeters, François Chaumont, Catherine Navarre
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引用次数: 0

Abstract

Nicotiana tabacum BY-2 cell suspension cultures are a powerful platform for producing recombinant glycoproteins such as immunoglobulins. Extensive efforts have been devoted to engineering the N-glycosylation pathway of BY-2 cells to overcome differences between mammalian and plant N-glycans. However, the mucin-type O-glycosylation pathway is absent in plant cells. This modification, which consists of glycan attachment to serine or threonine residues, is important in many human proteins, but is also highly complex. In this regard, plants offer a unique opportunity to engineer this pathway de novo without the interplay of many competing enzymes. In this study, transgenic BY-2 cell lines expressing the enzymes responsible for the formation of Core 1 and Tn antigen glycans were generated. First, GalNAc-O-glycosylation was initiated by the expression of human GalNAcT2. This O-glycan was then elongated by co-expression of Drosophila C1GalT1 to form the core 1 structure. Human IgA1 was produced in these engineered BY-2 cell lines and the presence of mucin-type O-glycans was confirmed by lectin blotting. The precise O-glycosylation profile of the hinge region was determined by mass spectrometry and showed the almost complete disappearance of pentoses and the presence of core 1 O-glycans.

在烟草BY-2细胞中产生具有明确黏液型o -聚糖的重组IgA1。
烟草BY-2细胞悬浮培养是生产重组糖蛋白(如免疫球蛋白)的强大平台。为了克服哺乳动物和植物n -聚糖之间的差异,人们已经投入了大量的努力来设计BY-2细胞的n -糖基化途径。然而,在植物细胞中不存在粘蛋白型o糖基化途径。这种由聚糖附着在丝氨酸或苏氨酸残基上的修饰在许多人类蛋白质中都很重要,但也非常复杂。在这方面,植物提供了一个独特的机会来重新设计这一途径,而不需要许多竞争酶的相互作用。在本研究中,产生了表达负责形成Core 1和Tn抗原聚糖的酶的转基因BY-2细胞系。首先,galnac - o糖基化是由人GalNAcT2的表达引发的。然后,这个o-聚糖通过果蝇C1GalT1的共表达被拉长,形成核心1结构。人IgA1在这些工程by -2细胞系中产生,凝集素印迹证实了黏液型o -聚糖的存在。质谱测定了铰链区精确的o -糖基化谱,显示戊糖几乎完全消失,核心的1 - o -聚糖存在。
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来源期刊
Frontiers in Plant Science
Frontiers in Plant Science PLANT SCIENCES-
CiteScore
7.30
自引率
14.30%
发文量
4844
审稿时长
14 weeks
期刊介绍: In an ever changing world, plant science is of the utmost importance for securing the future well-being of humankind. Plants provide oxygen, food, feed, fibers, and building materials. In addition, they are a diverse source of industrial and pharmaceutical chemicals. Plants are centrally important to the health of ecosystems, and their understanding is critical for learning how to manage and maintain a sustainable biosphere. Plant science is extremely interdisciplinary, reaching from agricultural science to paleobotany, and molecular physiology to ecology. It uses the latest developments in computer science, optics, molecular biology and genomics to address challenges in model systems, agricultural crops, and ecosystems. Plant science research inquires into the form, function, development, diversity, reproduction, evolution and uses of both higher and lower plants and their interactions with other organisms throughout the biosphere. Frontiers in Plant Science welcomes outstanding contributions in any field of plant science from basic to applied research, from organismal to molecular studies, from single plant analysis to studies of populations and whole ecosystems, and from molecular to biophysical to computational approaches. Frontiers in Plant Science publishes articles on the most outstanding discoveries across a wide research spectrum of Plant Science. The mission of Frontiers in Plant Science is to bring all relevant Plant Science areas together on a single platform.
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