ISSN:
0006-3592
Keywords:
specific secretion rate
;
animal cell culture
;
hybridoma
;
osmotic stress
;
variance of specific secretion rate
;
Chemistry
;
Biochemistry and Biotechnology
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Biology
,
Process Engineering, Biotechnology, Nutrition Technology
Notes:
The specific secretion rate (q, μg protein secreted/viable cell-h) and its variance are very useful to compare the capability of cell lines for protein secretion. An assessment of specific secretion rate variability is also beneficial and important when the specific secretion rate is to be used as an on-line process parameter to monitor culture production behavior or for in-process decisionmaking. Experimental errors in mammalian cell culture (e.g., protein concentration measurement and cell counting) and estimation error in the method of calculating q contribute to the total variance of the specific secretion rate. Although the variance of q is essential for comparing the differences between cell lines and the response of the same cell line to different nutrient or environmental conditions, few methods for calculating the variance of the specific secretion rate have been reported. As a model system, we have used the weighted jackknife method and the delta method to calculate the variance in the specific secretion rate of a murine monoclonal antibody (qmAb) determined by a differential method. These methods were applied to calculate qmAb and its standard deviation to determine the change in qmAb kinetics during batch culture of the 9.2.27 hybridoma in response to growth in hyperosmotic media or osmotic stress. Without osmotic stress, during exponential growth in DMEM + 5% FBS spinner culture, the estimate of qmAb decreases at least threefold. Results indicate that the 9.2.27 hybridoma responds to hyperosmotic media (400 mOsm, 470 mOsm) by significantly reducing the degree of qmAb decrease in the exponential phase, thus maintaining a higher qmAb through the stationary phase. The trend of qmAb during the batch cultures studied is further confirmed by t-test. Osmotic stress is statistically shown to be able to alter significantly the hybridoma-specific mAb secretion kinetics during batch culture. Determination of the variance of specific secretion rate using the weighted jackknife method offers a powerful approach for establishing the confidence limits of specific protein secretion rate between cell cultures in different nutritional or osmotic environments. © 1996 John Wiley & Sons, Inc.
Additional Material:
7 Ill.
Type of Medium:
Electronic Resource
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