Indirect 125I-labeled protein A assay for monoclonal antibodies to cell surface antigens
Journal of Immunological MethodsPublished 1 December 1979
Joseph P. Brown, John Tamerius, Ingegerd Hellström
Citations83
SJR quartileQ3
SJR score0.53
SNIP0.53
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TL;DR
The assay is used to select hybridomas producing monoclonal antibodies to surface antigens of human melanomas and mouse sarcomas and large numbers of samples can be screened rapidly and inexpensively.
Abstract
An assay for detection of monoclonal hybridoma antibodies against cell surface antigens is described. Samples of spent medium from the hybridoma cultures are incubated in microtest wells with cells, either as adherent monolayers or in suspension. Antibodies bound to surface antigens are detected by successive incubations with rabbit anti-immunoglobulin serum and 125I-labeled protein A from Staphylococcus aureus, followed by autoradiography of the microtest plate or scintillation counting of the individual wells. Particular advantages of this assay for screening hybridomas are: (1) commerically available reagents are used, (2) antibodies of any species and of any immunoglobulin class or subclass can be detected, and (3) large numbers of samples can be screened rapidly and inexpensively. We have used the assay to select hybridomas producing monoclonal antibodies to surface antigens of human melanomas and mouse sarcomas.
Keywords
Medicine
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