Kushida A, Koga M, Takagi J, Saito Y
Department of Biological Sciences, Tokyo Institute of Technology, Yokohama, Japan.
Hybridoma. 1998 Apr;17(2):209-13. doi: 10.1089/hyb.1998.17.209.
We have developed a novel method for screening hybridoma clones producing antibodies against plasma membrane-associated materials. The method is based on the binding of cells by antigen-antibody complex formation to immobilized antibodies. The monoclonal antibodies (MAbs) were trapped by goat anti-mouse immunoglobulin antibodies, which were coated on the surface of 96-well plastic plates. Cells to be tested were then added to these plates and the number of bound cells was quantified by the activity of phosphatase of bound cells. This method allows quick and reliable screening of hybridoma clones without resorting to the use of expensive instruments such as a flow cytometer. We immunized mice with two different kinds of cells and obtained two interesting but contrasting MAbs reacting to cell-surface antigens: one interacts quite specifically with neuroblastoma cells and the other reacts equally with various cells with the exception of GOTO, a neuroblastoma cell.
我们开发了一种用于筛选产生针对质膜相关物质抗体的杂交瘤克隆的新方法。该方法基于抗原 - 抗体复合物形成使细胞与固定化抗体结合。单克隆抗体（MAbs）被包被在96孔塑料板表面的山羊抗小鼠免疫球蛋白抗体捕获。然后将待测细胞添加到这些板中，并通过结合细胞的磷酸酶活性对结合细胞的数量进行定量。这种方法无需使用诸如流式细胞仪等昂贵仪器，就能快速且可靠地筛选杂交瘤克隆。我们用两种不同类型的细胞免疫小鼠，获得了两种有趣但截然不同的与细胞表面抗原反应的单克隆抗体：一种与神经母细胞瘤细胞特异性相互作用，另一种除了神经母细胞瘤细胞GOTO外，与各种细胞均有同等反应。
