Development of a quantitative cell-based intracellular ELISA for the screening of B cell hybridoma supernatants: a novel rapid assay to detect positive clones.
The primary screening of hybridoma clones secreting monoclonal antibodies (MAbs) requires the testing of a large number of hybridoma culture supernatants within a short time and is very labor-intensive. In addition, the type of antigen and its location in the cell have to be considered when selecting the appropriate screening procedure, but relatively few reagents are available for analyzing these molecules. We have developed an intracellular and cell surface ELISA technique for screening hybridoma supernatants that hastens the screening procedure of a large number of clones in a short period of time, as the supernatants of fused cells grown in 96-well plates are used directly in the assay. This novel screening technique is rapid, sensitive, specific, and applicable to MAbs specific for a wide variety of intracellular and/or cell surface proteins.
Renukaradhya
J Gourapura
JG
Department of Microbiology and Immunology, Indiana University School of Medicine, Walther Oncology Center, and the Walther Cancer Institute, Indianapolis, Indiana 46202-5181, USA.
Sriram
Venkataraman
V
Polakova
Katarina
K
Russ
Gustav
G
Brutkiewicz
Randy R
RR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D018949:Antigens, CD1 / Q000276:immunology; D055610:Antigens, CD1d; D001402:B-Lymphocytes / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005455:Fluorescent Antibody Technique; D006825:Hybridomas / Q000276:immunology*; D008297:Male; D051379:Mice
D000911:Antibodies, Monoclonal; D018949:Antigens, CD1; D055610:Antigens, CD1d