Abstract
Publisher Summary This chapter describes the steps involved in estimation of biomass in growing cell lines by luminometric ATP assay. Methods for studying the reliability of the results are described. In particular, estimation of energy charge (EC) by a new method for assay of ATP, ADP, and AMP was found to be valuable. The activity of several adenosine nucleotide converting enzymes as a function of the energy charge—that is, (ATP + 0.5 ADP) / (ATP + ADP + AMP)—indicates that the energy charge in intact metabolizing cells should be stabilized in the interval 0.8–0.9. By measuring all three adenine nucleotides, the measure of the physiological status of the cells is obtained that provides information on whether ATP can be used to estimate biomass or if the energy metabolism is disturbed. By the method described in the chapter, all three adenine nucleotides are determined in a single aliquot from the cell extract. Measurement of energy charge gives a reliable estimation of the accuracy of using ATP as a biomass parameter. Growth monitoring by assay of adenine nucleotides includes the following steps: (1) cultivation of cells, (2) sampling and pretreatment of samples, (3) extraction of adenine nucleotides from cells, (4) luminometric assay of adenine nucleotides, and (5) correlation to other biomass estimations.
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