PubMed1983
A new system for the screening of hybrid cultures in high number for possible production/secretion of antibodies (immunoglobulin) is presented. In the hybridoma technique for the production of monoclonal antibodies, myeloma cells are fused with B-lymphoblasts. This fusion is induced and often polyethylene glycol is used for this. Soon after fusion, a multitude of small colonies appears. Because of the biochemical characteristics of the myeloma cells, a selection, which only leaves fused cells, is possible. Among these colonies of fused cells, some will be antibody producing and secreting, and some of these will have the correct specificity. This specificity is towards the antigen originally used for immunization of the animal (often mouse). Because of the high number of fused cells, it is important to eliminate competition among different clones, wherefore seeding is done in microtest plates. Even so, more fused cells can end up in the same well, and competition can occur. Therefore, it is important to identify the clones secreting antibody of correct specificity as soon as possible. In the systems employed until now, medium had to be removed from the culture microtest plates. This induces a change in composition with replenishment, and so is not desired too often. Furthermore, this pipetting of small amounts of medium is laborious having 5, 10, or more plates. Therefore, a new system has been developed, which does not remove any medium, that has EIA sensitivity, is fast, and with very few necessary pipettings. The system uses a transferred solid phase, and is called NUNC-TSP-Screening System.