Rapid quality evaluation of hybridomas using ELISPOT and cell-ELISA techniques
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1University of Bergen, Norway.
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.
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.