用抗多糖单克隆抗体分析大鼠肠黏膜中位点依赖性粘蛋白。

IF 3.1 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Makoto Kurihara, Yukinobu Goso, Rei Kawashima, Takafumi Ichikawa, Kazuhiko Ishihara
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引用次数: 0

摘要

肠凝胶形成粘蛋白是一种高分子量的糖蛋白,其聚糖部分根据肠道部位的不同而变化。虽然分析粘蛋白聚糖的手段有限,但单克隆抗体有望成为一种强大的分析工具。本研究以大鼠肠粘蛋白为免疫原,制备了12种单克隆抗体,并对抗体的表位进行了研究。用与血型相关抗原的聚糖和粘蛋白聚糖合成的新糖脂的树脂分析表明,大多数抗体识别聚糖。此外,一些在聚糖上识别血型相关抗原,而另一些识别不成熟合成的粘蛋白型聚糖。用抗体对大鼠空肠、回肠、近端结肠和远端结肠进行免疫染色,产生位点依赖的肠道图像。采用抗体偶联柱亲和层析法分离大鼠小肠粘蛋白。产生了具有不同聚糖组成的粘蛋白,表明获得位点特异性粘蛋白的可能性。具有不同聚糖组成的位点特异性粘蛋白可产生位点特异性聚糖功能,本研究开发的抗体可作为分析位点特异性粘蛋白的有用工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Analysis of site-dependent mucins in rat intestinal mucosa using anti-glycan monoclonal antibodies.

Intestinal gel-forming mucins are high-molecular weight glycoproteins, and their glycan moieties vary depending on the intestinal site. Although the means of analyzing mucin glycans are limited, monoclonal antibodies are expected to be a powerful analysis tool. In this study, 12 monoclonal antibodies were generated using rat intestinal mucin as the immunogen, and the epitopes of the antibodies were studied. The analyses using resins with the glycans of blood group-associated antigens and neoglycolipids synthesized from mucin glycans revealed that most antibodies recognize glycans. Furthermore, some recognize blood group-related antigens on glycans, whereas others recognize immaturely synthesized mucin-type glycans. Immunostaining of the rat jejunum, ileum, proximal colon, and distal colon with antibodies produced a site-dependent staining intestinal image. Affinity chromatography using an antibody-conjugated column was utilized to fractionate rat small intestinal mucins. Mucins with various glycan compositions were produced, suggesting the possibility of obtaining site-specific mucins. The site-specific mucins with various glycan compositions result in site-specific glycan functions, and the antibodies developed in this study could be useful tools for their analysis.

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来源期刊
Glycoconjugate Journal
Glycoconjugate Journal 生物-生化与分子生物学
CiteScore
6.00
自引率
3.30%
发文量
63
审稿时长
1 months
期刊介绍: Glycoconjugate Journal publishes articles and reviews on all areas concerned with: function, composition, structure, biosynthesis, degradation, interactions, recognition and chemo-enzymatic synthesis of glycoconjugates (glycoproteins, glycolipids, oligosaccharides, polysaccharides and proteoglycans), biochemistry, molecular biology, biotechnology, immunology and cell biology of glycoconjugates, aspects related to disease processes (immunological, inflammatory, arthritic infections, metabolic disorders, malignancy, neurological disorders), structural and functional glycomics, glycoimmunology, glycovaccines, organic synthesis of glycoconjugates and the development of methodologies if biologically relevant, glycosylation changes in disease if focused on either the discovery of a novel disease marker or the improved understanding of some basic pathological mechanism, articles on the effects of toxicological agents (alcohol, tobacco, narcotics, environmental agents) on glycosylation, and the use of glycotherapeutics. Glycoconjugate Journal is the official journal of the International Glycoconjugate Organization, which is responsible for organizing the biennial International Symposia on Glycoconjugates.
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