Tanaka T, Kurth R
J Immunol Methods. 1984 Dec 31;75(2):217-25. doi: 10.1016/0022-1759(84)90105-4.
A small scale solid-phase radioimmunoassay employing 125I-labelled protein A is described which is suitable for screening large numbers of monoclonal antibodies directed against antigens which can be prepared in small amounts only, for example oncoviral proteins. The use of polystyrene Terasaki microtest plates instead of 96-well microtitre plates reduces the amount of antigen required for screening hybridoma supernatants to less than 30 ng/well. This method facilitates washing procedures and reduces the quantity of radioactive waste. The sensitivity and specificity of the method is demonstrated by the isolation and initial characterization of monoclonal antibodies specific for avian oncoviral transforming, structural and polymerase proteins.
本文描述了一种使用¹²⁵I标记蛋白A的小规模固相放射免疫测定法,该方法适用于筛选大量针对只能少量制备的抗原(如肿瘤病毒蛋白)的单克隆抗体。使用聚苯乙烯寺崎微量滴定板而非96孔微量滴定板,可将筛选杂交瘤上清液所需的抗原量减少至每孔不到30 ng。该方法便于洗涤操作并减少放射性废物量。通过分离和初步鉴定针对禽肿瘤病毒转化蛋白、结构蛋白和聚合酶蛋白的单克隆抗体,证明了该方法的敏感性和特异性。