ISSN:
1573-0778
Schlagwort(e):
CHO cells
;
gel microdrops
;
human antibody
;
population parameters
;
productivity
;
stability
Quelle:
Springer Online Journal Archives 1860-2000
Thema:
Biologie
,
Medizin
,
Werkstoffwissenschaften, Fertigungsverfahren, Fertigung
Notizen:
Abstract The long-term stability of high-level expression is the mostimportant factor to consider when choosing cell lines for the expression of recombinant proteins. Declining volumetricyields in large-scale fermentation can be caused by changes affecting the cell population as a whole such as loss in viability, depletion of nutrients or accumulation of metabolites affecting cell growth. Alternatively, geneticinstability may lead to the outgrowth of a less productive,metabolically favored sub-population. Currently a variety ofparameters are measured to monitor the condition of cells infermenters including glucose uptake, lactate accumulation andoxygen consumption; in addition, periodic viable cell countsallow the determination of the growth rate and viability of the population. All of these methods measure the condition ofthe cell population as a whole and changes must involve a significantly large proportion of the total culture in orderto be detectable. Here we report on a method that allows theevaluation of the productivity of individual cells. Using the gel microdrop secretion assay, we detected the appearance ofa sub-population of cells with lower productivity. Subsequentanalysis of the culture confirmed the existence of lower productivity cells with a lower vector copy number. Therefore,the single cell secretion assay proved to be a rapid method todetect and isolate a low productivity variant of the producer cell line.
Materialart:
Digitale Medien
URL:
http://dx.doi.org/10.1023/A:1008186113245
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