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The optimisation of immunoglobulin secretion in vitro by mouse spleen cells and hybridoma cells
1International Laboratory for Research on Animal Diseases, Nairobi, Kenya.
Optimal culture conditions and cell densities for antibody production by spleen cells from mice infected with Trypanosoma brucei, and by hybridoma cells, were assessed in vitro by a silver-immunogold (SIG) blot technique and an enzyme-linked immunosorbent assay (ELISA). The blot assay measured the proportion of secreting cells in the population and the ELISA measured the quantity of synthesised and secreted immunoglobulin (Ig) in cell lysates and in 3-h culture supernatants. Marked inhibition of Ig secretion occurred when cells were cultured in conditions of confluence. When cells were maintained in rotating cultures, no inhibition was seen. These cells maintained a steady rate of Ig synthesis and secretion over 3 h.
Optimal culture conditions and cell densities for antibody production by spleen cells from mice infected with Trypanosoma brucei, and by hybridoma cells, were assessed in vitro by a silver-immunogold (SIG) blot technique and an enzyme-linked immunosorbent assay (ELISA). The blot assay measured the proportion of secreting cells in the population and the ELISA measured the quantity of synthesised and secreted immunoglobulin (Ig) in cell lysates and in 3-h culture supernatants. Marked inhibition of Ig secretion occurred when cells were cultured in conditions of confluence. When cells were maintained in rotating cultures, no inhibition was seen. These cells maintained a steady rate of Ig synthesis and secretion over 3 h.