Rapid quality evaluation of hybridomas using ELISPOT and cell-ELISA techniques.
We have used the ELISPOT assay in combination with ELISA procedures for rapid evaluation of properties of three different murine hybridoma cell lines, 104-I, -B and -G, secreting IgG1 kappa monoclonal antibodies against a 104-mer synthetic peptide from the C-terminal part of HIV-1 p24. By conventional ELISA we obtained data suggesting that the three monoclonal antibodies had different affinities. By cell-ELISA we found that the IgG1 kappa secretion rate varied between cells (4,000 to 14,000 antibody molecules/cell/min), and ELISPOT showed that only 4-5% of 104-I cells gave antigen-specific spots, indicating a cell population with diverse properties. We recommend that the ELISPOT and cell-ELISA techniques should be used routinely to supplement conventional ELISA procedures for rapid evaluation of hybridoma properties.
Haaheim
L R
LR
University of Bergen, Norway.
Jonsson
R
R
D000818:Animals; D000915:Antibody Affinity; D002460:Cell Line; D002999:Clone Cells; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D016655:HIV Core Protein p24 / Q000276:immunology*; D006825:Hybridomas / Q000276:immunology*; D007074:Immunoglobulin G / Q000096:biosynthesis*; D051379:Mice
D016655:HIV Core Protein p24; D007074:Immunoglobulin G