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首页> 外文会议>International Wood Biotechnology Symposium (IWBS) Mar 14-17, 2001, Narita, Chiba, Japan >ISOLATION OF MONOCLONAL ANTIBODIES RECOGNIZING XYLEM CELL WALL COMPONENTS BY USING A PHAGE DISPLAY SUBTRACTION METHOD
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ISOLATION OF MONOCLONAL ANTIBODIES RECOGNIZING XYLEM CELL WALL COMPONENTS BY USING A PHAGE DISPLAY SUBTRACTION METHOD

机译：噬菌体色谱分离法分离识别木聚糖壁成分的单克隆抗体。

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摘要

Monoclonal antibodies (mAbs) recognizing differentiated cell-specific wall components are useful in distinguishing types of cells and dissecting developmental processes. We tried to generate such mAbs using a strategy consisting of (ⅰ) isolation of cell walls from synchronously differentiating cells of an in vitro Zinnia culture system, which reflects vascular differentiation in plants (ⅱ) construction of phage display library of recombinant antibody against the cell walls (ⅲ) screening via biopannings with subtractive procedures using non-differentiating cell walls. The advantage of this strategy is that purposive mAbs can be easily screened without purifying antigens of interest. Moreover, this approach can be applied to search for novel subpopulations of cells that are difficult to address by conventional methods because of small subpopulations in heterogeneous mixtures. As a result, we succeeded in isolation of three mAbs, designated CN 8, XD 3, and XD 27. Immunolocalization analyses in Zinnia plants revealed that CN 8 epitope localized in walls of immature tracheary elements (TEs) and xylem parenchyma cells, XD 3 epitope localized in walls of TE precursors and immature fiber cells, and XD 27 epitope localized only in walls of immature TEs. In the Zinnia culture system, these three mAbs distinguished subpopulations of cells in different developmental stages. These results demonstrate that some wall components change dynamically in association with xylem cell differentiation, as cell-surface antigens of animal cells. These mAbs, therefore, are useful as molecular markers to dissect the vascular developmental stage and also as tools to isolate specific xylem cells using a cell sorter.
机译：识别分化的细胞特异性壁成分的单克隆抗体（mAb）可用于区分细胞类型和解剖发育过程。我们尝试使用以下策略来生成此类单克隆抗体：（ⅰ）从体外百日草培养系统的同步分化细胞中分离细胞壁，从而反映出植物中的血管分化（ⅱ）构建针对该细胞的重组抗体的噬菌体展示文库使用非分化细胞壁通过消减程序通过生物淘选筛选细胞壁（）。该策略的优势在于，无需纯化目标抗原即可轻松筛选目的性mAb。而且，该方法可用于搜索由于异质混合物中的小亚群而难以通过常规方法处理的细胞的新亚群。结果，我们成功分离了三个单克隆抗体，分别命名为CN 8，XD 3和XD27。百日草植物中的免疫定位分析表明，CN 8表位位于未成熟气管元件（TEs）和木质部薄壁细胞XD 3的壁中。抗原决定簇位于TE前体和未成熟纤维细胞的壁中，而XD 27抗原决定簇仅位于未成熟TE的壁中。在百日菊属培养系统中，这三个mAb区分了处于不同发育阶段的细胞亚群。这些结果表明，一些壁成分与木质部细胞分化相关地动态变化，作为动物细胞的细胞表面抗原。因此，这些mAb可用作解剖血管发育阶段的分子标记，也可用作使用细胞分选仪分离特定木质部细胞的工具。

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