Screening of Hybridoma Clones Producing Antibodies Against Plasma Membrane-Associated Materials

Abstract

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—ntibody 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.