TY - JOUR
T1 - A semi-automated fluorescent (SAF) assay using viable, whole cells for screening hybridoma supernatants
AU - Liebert, M.
AU - Laino, L.
AU - Wahl, R. L.
N1 - Funding Information:
One of the most time-consuming and labor-intensive steps in the production of monoclonal antibodies is the screening. Screening is a critical step in the development of monoclonal antibodies of desired specificity, because screening results are used to determine which cells will be subcloned and saved. Time is also a limiting factor because of cell growth while waiting for results. Therefore, the assay chosen must be accurate, fast, sensitive, Correspondence to: R.L. Wahl, Division of Nuclear Medicine, University of Michigan Medical Center, 1500 Medical Center Drive, Box 0028, Ann Arbor, MI 48109-0028, U.S.A. * Supported in part by PHS R01-CA40497-02, R01-CA41531-02, R01-CA33802 awarded by NCI, DHHS, and by DOE Contract DE-AC02-76EV002031.
PY - 1987/6/16
Y1 - 1987/6/16
N2 - 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.
AB - 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.
KW - Hybridoma screening procedure
KW - Immunofluorescence
KW - Monoclonal antibody
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U2 - 10.1016/0022-1759(87)90220-1
DO - 10.1016/0022-1759(87)90220-1
M3 - Article
C2 - 3302046
AN - SCOPUS:0023218431
SN - 0022-1759
VL - 101
SP - 85
EP - 90
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
IS - 1
ER -