Animal Cells as Bioreactors by Terence Cartwright

By Terence Cartwright

This booklet covers all elements of the hot applied sciences had to flip animal cells into a suitable and low in cost software for drug construction. This comprises editing them genetically so they produce the best product in excessive yield, getting them to develop reproducibly on an commercial scale, and extracting the mandatory product from them. It additionally covers organic questions of safety, and the verification of the chemical and organic nature of the protein drug produced. The paintings covers the newest advancements in all of those parts and the way all of them have to be built-in for the layout of an efficient biotechnological construction procedure.

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In a subsequent study (Bebbington et al, 1992) the successful use of the GS amplification system in NSO myeloma cells was reported. As previously discussed, vector amplification in myeloma cells using the DHFR system (Okamoto et al, 1990) has been achieved in some cases, but in the presence of endogenous DHFR activity, very high levels of MTX were required to obtain satisfactory amplification. In other studies, attempts to amplify antibody genes in myeloma cells using MTX did not produce significant amplification (Gillies et al, 1989).

Thus, optimal expression was obtained using a defined glutamine-free medium which had to be supplemented with additional glutamate, asparagine and nucleotides to compensate for the absence of glutamine. The asparagine requirement of NSO cells is unusual and possibly was needed as a metabolic source of ammonia. The development of a suitable amino acid fed-batch regime also increased yield significantly and air-lift fermentation was shown to be superior to fermentation in stirred tanks. In other studies it has also been reported that methionine supplementation of the medium may be useful because MSX is partially detoxified through the methionine catabolic pathway and excess methionine can saturate this pathway, with a consequent sparing effect on MSX and thus effectively enhance its toxicity (Bebbington and Hentschel, 1987).

Considerable success in maintaining quality and in limiting batch-to-batch variation has been achieved. However, despite all the efforts to standardize conditions of collection, foetal bovine serum remains a by-product of the dairy industry whose availability is determined by current dairy policies which in turn determine the number of pregnant cows going to slaughter. For example, foetal bovine serum availability increased suddenly in 1986 when large numbers of dairy cows were slaughtered in the United States due to a revision of the structure of dairy subsidies.

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