Sammons D W, Sanchez E, Darlington G J
In Vitro. 1980 Nov;16(11):918-24. doi: 10.1007/BF02619329.
A screening technique was developed for the identification of clones of hepatoma cells that secrete albumin. The technique employs the overlay of a 1% agarose solution containing antiserum to albumin onto clones of hepatoma cells. A distinct immunoprecipitation complex is formed in the immuno-overlay that corresponds directly to the position of each secreting clone. Clones deficient in albumin secretion do not form an immunoprecipitate. Thus comparison of the immuno-overlay and the cell colonies results in identification of variant clones as well as those capable of secretion. Biochemical characterization of the region of agarose overlay from secreting and nonsecreting clones demonstrates the specificity of the method and its potential for selection of colonies that are secreting other hepatic or cellular proteins.
开发了一种用于鉴定分泌白蛋白的肝癌细胞克隆的筛选技术。该技术是将含有抗白蛋白血清的1%琼脂糖溶液覆盖在肝癌细胞克隆上。在免疫覆盖物中形成一个明显的免疫沉淀复合物,该复合物直接对应于每个分泌克隆的位置。缺乏白蛋白分泌的克隆不会形成免疫沉淀物。因此,比较免疫覆盖物和细胞集落可鉴定出变异克隆以及能够分泌的克隆。对分泌克隆和非分泌克隆的琼脂糖覆盖区域进行生化特性分析,证明了该方法的特异性及其在选择分泌其他肝脏或细胞蛋白的集落方面的潜力。