Evidence for posttranscriptional stimulation of monoclonal antibody secretion by L-glutamine during slow hybridoma growth.

The addition of 5-40 mM L-glutamine to batch cultures of a murine hybridoma following the cessation of rapid growth significantly stimulated monoclonal antibody (mAb) synthesis and secretion per cell. Stimulation of mAb secretion following the cessation of rapid growth was also observed in response to addition of mitochondrial intermediates of glutamate oxidation and was not found to be the result of release of transiently stored mAb. Less than 1% of the secreted mAb was detected by ELISA in isolated hybridoma lysosomes. This stimulation was posttranscriptional and not the result of enhancement of levels of mAb mRNAs or stabilization of heavy (H) or light (L) chain encoding message. Sub-inhibitory levels of lysosomotrophic weak bases stimulated release of lysosomal contents but did not result in release of intact or partially degraded mAb. Inhibition of aspartic proteinase activity secreted by the hybridoma did not enhance mAb secretion even though a high level of mAb degrading proteinase activity was continuously secreted during both rapid and slow growth. These responses indicate that during slow growth, the addition of L-glutamine increases the availability of cellular ATP generated by mitochondrial respiration which stimulates some posttranscriptional step in the pathway of mAb secretion such as the rate of H or L chain translation, chain assembly, interorganelle transport or vesicular transport from the Golgi to the cell membrane.