Defined protein and animal component-free NS0 fed-batch culture
2007 (English)In: Biotechnology and Bioengineering, ISSN 0006-3592, E-ISSN 1097-0290, Vol. 98, no 6, 1183-1194 p.Article in journal (Refereed) Published
A chemically defined protein and animal component-free fed-batch process for an NS0 cell line producing a human IgG(1) antibody has been developed. The fed-batch feed profile was optimised in a stepwise manner. Depletion of measurable compounds was determined by direct analysis. The cellular need for nonmeasurable compounds was tested by continued culturing of cell suspension, removed from the bioreactor, in shake-flasks supplemented with critical substances. In the final fed-batch culture, 8.4 x 10(6) viable cells mL(-1) and 625 mg antibody L-1 was obtained as compared to 2.3 X 10(6) cells mL(-1) and 70 mg antibody L-1 in batch. The increase in cell density, in combination with a prolonged declining phase where antibody formation continued, resulted in a 6-fold increase in total cell yield, a 10.5-fold increase 6.2 in viable cell hours and an 11.4-fold increase in product yield. These improvements were obtained by using a feed with glucose, glutamine, amino acids, lipids, sodium selenite, ethanolamine and vitamins. Specifically, supplementation with lipids (cholesterol) had a drastic effect on the maximum viable cell density. Calcium, magnesium and potassium were not depleted and a feed also containing iron, lithium, manganese, phosphorous and zinc did not i significantly enhance the cell yield. The growth and death profiles in the final fed-batch indicated that nutrient deprivation was not the main cause of cell death. The ammonium concentration and the osmolality increased to potentially inhibitory levels, but an imbalance in the supply of growth/survival factors may also contribute to termination of the culture.
Place, publisher, year, edition, pages
2007. Vol. 98, no 6, 1183-1194 p.
NSO cells, protein-free medium, fed-batch culture, cholesterol, vitamins
IdentifiersURN: urn:nbn:se:kth:diva-6314DOI: 10.1002/bit.21509ISI: 000250813200007ScopusID: 2-s2.0-36749087591OAI: oai:DiVA.org:kth-6314DiVA: diva2:10994
QC 20100907. Uppdaterad från Submitted till Published (20100907)2006-11-012006-11-012010-09-07Bibliographically approved