牛颞下颌关节盘硫酸大分子软骨素蛋白多糖单克隆抗体

T. Nakano , S. Imai , T. Koga , C.M. Dodd , P.G. Scott
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引用次数: 26

摘要

用从牛颞下颌关节盘分离纯化的硫酸软骨素蛋白多糖免疫小鼠,获得了4个产生单克隆抗体的杂交细胞系(AC2、AH12、DB10和DD11)。通过酶联免疫吸附试验和完整蛋白聚糖和由糖苷酶和蛋白酶制成的酶切物的免疫印迹染色模式对这些表位进行了部分表征。所有四种单克隆抗体似乎都能识别某种形式的硫酸角蛋白,尽管其中两种(AC2和DD11)的表位可能相同。一种抗体(AH12)对角膜硫酸角蛋白几乎没有反应性,但除了对大的硫酸软骨素蛋白聚糖外,还对从角膜盘中提取的小的硫酸角蛋白聚糖进行了染色。这些抗体用于椎间盘切片的免疫组织化学染色,显示与大硫酸软骨素蛋白多糖相关的硫酸角蛋白集中在结构的内部和远离外围,但靠近上下表面,这种模式可能反映了细胞外基质对咀嚼施加在其上的机械应力的适应。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Monoclonal Antibodies to the Large Chondroitin Sulphate Proteoglycan from Bovine Temporomandibular Joint Disc

Four hybrid cell lines producing monoclonal antibodies (designated AC2, AH12, DB10 and DD11) were derived from mice immunized with the large chondroitin sulphate proteoglycan isolated and purified from the bovine temporomandibular joint disc. The epitopes were partially characterized by enzyme-linked immunosorbent assays and staining patterns on immunoblots of intact proteoglycans and digests made with glycosidases and proteinases. All four monoclonal antibodies appeared to recognize some form of keratan sulphate although the epitopes for two (AC2 and DD11) were probably identical. One antibody (AH12) showed almost no reactivity with corneal keratan sulphate but stained a small keratan sulphate proteoglycan extracted from the disc, in addition to the large chondroitin sulphate proteoglycan. These antibodies were used for immunohistochemical staining of sections of the disc and showed that keratan sulphate associated with the large chondroitin sulphate proteoglycan was concentrated inside and away from the periphery of the structure but close to the inferior and superior surfaces, in a pattern which may reflect the adaptation of the extracellular matrix to the mechanical stresses placed on it by mastication.

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