Funderburgh J L, Stenzel-Johnson P R, Chandler J W
Curr Eye Res. 1982;2(11):769-76. doi: 10.3109/02713688209020010.
Six mouse hybrid cell lines were isolated which secrete antibodies to rabbit corneal proteoglycan. All six antibodies interacted with the same fraction of the proteoglycan, precipitating approximately 50% of proteoglycan labelled in the protein moiety. A radioimmunoassay using these antibodies measured concentrations as low as 1 g/ml unlabelled rabbit corneal proteoglycan. Human corneal proteoglycan, corneal keratan sulfate, and an oligosaccharide fraction from corneal digests all interacted with the antibodies at concentrations similar to whole rabbit proteoglycan. Proteoglycans from cultured rabbit stromal fibroblasts and from sclera were 20 to 50-fold less effective in competition for antibody. Endo-beta-galactosidase treatment of proteoglycan reduced antibody binding, but protease or chondroitinase treatments did not. Labelled proteoglycan separated by antibody affinity chromatography contained only keratan sulfate, whereas proteoglycan not bound to affinity columns contained only chondroitin sulfate. The antibodies appear to recognise a carbohydrate structure found only on corneal keratan sulfate proteoglycan. This structure can serve as a basis for separation. This structure can serve as a basis for separation of corneal proteoglycan types using antibody affinity chromatography.
分离出六种小鼠杂交细胞系，它们分泌抗兔角膜蛋白聚糖的抗体。所有六种抗体都与蛋白聚糖的同一部分相互作用，沉淀出蛋白质部分中约50%的标记蛋白聚糖。使用这些抗体的放射免疫测定法可检测低至1μg/ml的未标记兔角膜蛋白聚糖浓度。人角膜蛋白聚糖、角膜硫酸角质素以及角膜消化物中的寡糖部分，在与全兔蛋白聚糖相似的浓度下均与抗体相互作用。来自培养的兔基质成纤维细胞和巩膜的蛋白聚糖在竞争抗体方面的效力要低20至50倍。蛋白聚糖经内切β-半乳糖苷酶处理后抗体结合减少，但蛋白酶或软骨素酶处理则不然。通过抗体亲和色谱法分离的标记蛋白聚糖仅含有硫酸角质素，而未结合到亲和柱上的蛋白聚糖仅含有硫酸软骨素。这些抗体似乎识别仅存在于角膜硫酸角质素蛋白聚糖上的碳水化合物结构。该结构可作为分离的基础。该结构可作为使用抗体亲和色谱法分离角膜蛋白聚糖类型的基础。
