A semi-automated fluorescent (SAF) assay using viable, whole cells for screening hybridoma supernatants.
In the production of monoclonal antibodies, a rapid, sensitive, accurate assay is needed for the critical step of screening. We report the modification of an assay using viable whole cells for screening hybridoma supernatants. The modified assay uses fluorescent second antibodies for detection and has been adapted to an instrument capable of automating a number of assay steps. The modified assay is compared to a dot radioimmunoassay developed and used in our laboratory. The fluorescence assay is highly sensitive but shows more background effect, especially in samples with high protein content, such as ascites. The automated fluorescence assay is very rapid, capable of completing an assay in less than 90 min, and can be performed with minimal operator involvement. The assay was performed successfully with several different antibodies and cell types. This screening procedure should be especially useful for laboratories with large numbers of fusions to evaluate.
Liebert
M
M
Laino
L
L
Wahl
R L
RL
D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000096:biosynthesis; D000918:Antibody Specificity; D001201:Ascites / Q000378:metabolism; D001331:Automation; D003470:Culture Media / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D006825:Hybridomas / Q000378:metabolism*; D008722:Methods; D011863:Radioimmunoassay / Q000379:methods; D013997:Time Factors
D000911:Antibodies, Monoclonal; D003470:Culture Media