Caterson B, Christner J E, Baker J R
J Biol Chem. 1983 Jul 25;258(14):8848-54.
Monoclonal antibodies were raised against proteoglycan core protein isolated after chondroitinase ABC digestion of human articular cartilage proteoglycan monomer. Characterization of one of the monoclonal antibodies (1/20/5-D-4) indicated that it specifically recognized an antigenic determinant in the polysaccharide structure of both corneal and skeletal keratan sulfate. Enzyme immunoassay analyses indicated that the mouse monoclonal IgG1 recognized keratan sulfate in native proteoglycan aggregate and proteoglycan monomer preparations isolated from hyaline cartilages of a wide variety of animal species (human, monkey, cow, sheep, chicken, and shark cartilage). The 1/20/5-D-4 monoclonal antibody did not recognize antigenic determinants on proteoglycan isolated from Swarm rat chondrosarcoma. This finding is consistent with several biochemical analyses showing the absence of keratan sulfate in proteoglycan synthesised by this tissue. A variety of substructures isolated after selective cleavage of bovine nasal cartilage proteoglycan (Heinegård, D., and Axelsson, J. (1977) J. Biol. Chem. 252, 1971-1979) were used as competing antigens in radioimmunoassays to characterize the specificity of the 1/20/5-D-4 immunoglobulin. Substructures derived from the keratan sulfate attachment region of the proteoglycan (keratan sulfate peptides) showed the strongest inhibition. Both corneal and skeletal keratan sulfate peptides as competing antigens in radioimmunoassays showed similar inhibition when compared on the basis of their glucosamine content. Therefore, the 1/20/5-D-4 monoclonal antibody appears to recognize a common determinant in their polysaccharide moieties. Chemical desulfation of the keratan sulfate reduced the antigenicity of the glycosaminoglycan. The antibody did not recognize determinants present in dermatan sulfate, heparin, heparin sulfate, or hyaluronic acid.
针对人关节软骨蛋白聚糖单体经软骨素酶ABC消化后分离得到的蛋白聚糖核心蛋白制备了单克隆抗体。对其中一种单克隆抗体（1/20/5-D-4）的特性分析表明，它能特异性识别角膜硫酸角质素和骨骼硫酸角质素多糖结构中的一个抗原决定簇。酶免疫分析表明，小鼠单克隆IgG1能识别从多种动物物种（人、猴、牛、羊、鸡和鲨鱼软骨）的透明软骨中分离得到的天然蛋白聚糖聚集体和蛋白聚糖单体制剂中的硫酸角质素。1/20/5-D-4单克隆抗体不能识别从斯旺大鼠软骨肉瘤中分离得到的蛋白聚糖上的抗原决定簇。这一发现与多项生化分析结果一致，这些分析表明该组织合成的蛋白聚糖中不存在硫酸角质素。在放射免疫分析中，将牛鼻软骨蛋白聚糖经选择性裂解后分离得到的多种亚结构（海内加德，D.，和阿克塞尔松，J.（1977年）《生物化学杂志》252卷，1971 - 1979页）用作竞争抗原，以表征1/20/5-D-4免疫球蛋白的特异性。源自蛋白聚糖硫酸角质素连接区域的亚结构（硫酸角质素肽）显示出最强的抑制作用。在放射免疫分析中，以硫酸角质素肽作为竞争抗原，基于其氨基葡萄糖含量进行比较时，角膜硫酸角质素肽和骨骼硫酸角质素肽表现出相似的抑制作用。因此，1/20/5-D-4单克隆抗体似乎能识别其多糖部分中的一个共同决定簇。硫酸角质素的化学脱硫降低了糖胺聚糖的抗原性。该抗体不能识别存在于硫酸皮肤素、肝素、硫酸乙酰肝素或透明质酸中的决定簇。
