Liebert M, Ballou B, Taylor R J, Reiland J M, Hakala T R
J Immunol Methods. 1985 Dec 17;85(1):97-104. doi: 10.1016/0022-1759(85)90277-7.
Membranes prepared from a variety of solid tissues were used as solid-phase antigens for ELISA or RIA after fixation onto polylysine-primed 96-well plates. The preservation of antigens in these membrane preparations was tested by reactivity in ELISA using 2 monoclonal antibodies: W6/32, which recognizes an HLA framework antigen (a protein antigen) and anti-SSEA-1, directed to a carbohydrate antigen carried on glycoproteins. Levels of antigen deposition and usefulness as solid-phase antigens were assessed for ELISA as compared to RIA. Coated plates may be frozen for many months with preservation of antigenic activity. This method is relatively simple, rapid, and is useful for preparation of tissue antigens for immunoassay, especially for screening monoclonal antibodies.
从各种实体组织制备的膜在固定到经聚赖氨酸预处理的96孔板上后,用作ELISA或RIA的固相抗原。使用两种单克隆抗体通过ELISA反应性测试这些膜制剂中抗原的保存情况:W6/32,识别HLA框架抗原(一种蛋白质抗原);抗SSEA-1,针对糖蛋白上携带的碳水化合物抗原。与RIA相比,评估了ELISA中抗原沉积水平和作为固相抗原的有用性。包被的板可冷冻数月,抗原活性得以保留。该方法相对简单、快速,可用于制备免疫测定用的组织抗原,尤其适用于筛选单克隆抗体。