Donoso L A, Folberg R, Edelberg K, Arbizo V, Atkinson B, Herlyn M
J Histochem Cytochem. 1985 Dec;33(12):1190-6. doi: 10.1177/33.12.3905953.
A screening method is described to select monoclonal antibodies (Mabs) that bind to ocular melanoma-associated antigens (MAAs) retained in formalin-fixed, paraffin-embedded tissue sections. Small sections of epithelioid or spindle-cell-type uveal melanomas were cut into 2 mm cubes and reembedded in one block. Microslides were cut from this block and used to screen hybridoma supernatant fluid. Using this screening method, three MAbs were selected from two separate fusions of mouse myeloma cells with spleen cells of mice immunized previously with either ocular melanoma cells obtained fresh at enucleation or cells of a cutaneous melanoma cell line. Although all three MAbs showed similar specificities, MAb8-1H showed the strongest immunohistochemical reaction and was studied further in detail. MAb8-1H bound to 91% (71/79) of the choroidal or ciliochoroidal melanomas tested, indicating a high prevalence of this antigen in uveal melanomas. The antigen defined by MAb8-1H was isolated, purified, and partially characterized as a 40,000-50,000 dalton, highly glycosylated protein rich in glycine, serine, and glutamic acid, as is typical of a mucin-type glycoprotein.
本文描述了一种筛选方法，用于选择能与福尔马林固定、石蜡包埋组织切片中保留的眼黑色素瘤相关抗原（MAA）结合的单克隆抗体（MAb）。将上皮样或梭形细胞型葡萄膜黑色素瘤的小切片切成2毫米的方块，重新包埋在一个蜡块中。从该蜡块上切取微切片，用于筛选杂交瘤上清液。使用这种筛选方法，从两例单独的小鼠骨髓瘤细胞与脾细胞的融合中筛选出了三种单克隆抗体，所用脾细胞来自先前用摘除眼球时新鲜获得的眼黑色素瘤细胞或皮肤黑色素瘤细胞系的细胞免疫过的小鼠。尽管所有三种单克隆抗体都表现出相似的特异性，但单克隆抗体8-1H显示出最强的免疫组织化学反应，并对其进行了进一步详细研究。单克隆抗体8-1H与91%（71/79）的测试脉络膜或睫状体脉络膜黑色素瘤结合，表明该抗原在葡萄膜黑色素瘤中具有较高的发生率。单克隆抗体8-1H所定义的抗原被分离、纯化，并部分鉴定为一种40,000-50,000道尔顿的高度糖基化蛋白，富含甘氨酸、丝氨酸和谷氨酸，这是粘蛋白型糖蛋白的典型特征。
