Production of tpa using eukaryotic n prokaryotic cells - Essay Example

The Production Of tPA using eukaryotic n prokaryotic cells The Production Of tPA using eukaryotic n prokaryotic cells Cell culture is the growing of prokaryotic or eukaryotic cells under controlled conditions. Mammalian cell culture has a wide application in biotechnology where they have been utilized for the production of vaccines, proteins, hormones, antibodies and enzymes. The challenge with large growth mammalian cell cultures is the cost. The major reason being their demand for a complex media.

Also, these cells are usually susceptible to contaminations resulting from microbial infections. From the case study, it is evident that production of recombinant tPA in CHO (mammalian) cells is less costly than in E. coli (prokaryotic). This is because of the high cost of the purification scheme required on the tPA produced from E. coli. That from the mammalian cells does not require lots of purifications because the cell environment and content is similar to that of the body cells (in vitro).

tPA produced in E. coli required more purification to acquire the required state because it is composed of many components that are naturally not found with tPA in vitro conditions. The contents produced together with tPA in mammalian cells are similar to those in vitro hence are not foreign materials resulting to a less purification process. The E. coli based production process utilizes approximately sixteen purification steps (downstream) as compared to the five purification steps in CHO. Since the purification process in mammalian cells is not that complex less operating labour and supervision is required compared to the E.

coli based. The CHO process utilizes nine percent of its total cost on operating labour while E. coli based utilizes twenty two percent. Also, the cost of acquiring the different purification units required for the many purification stages is higher in the E. coli based. Mammalian based process utilizes only twenty five percent of the total production cost on recovery (purification) compare to eighty eight percent on E. coli based. For the solubilization and renaturation step in the E. coli based production, approximately five tonnes of urea and twenty-six tonnes of guanidine are required.

The bacteria lack efficient secretion mechanisms for tPA, therefore, the proteins are deposited as inclusion bodies. For the recovery of the proteins, a renaturation procedure is required. Since the proteins in the bacteria are produced in low concentrations (about 2.4 mg per litre) the practicability of a renaturation based production process is limited. The E. coli based production does yield to larger quantities of products that may be more effective in vivo, but it is limited by the necessity for the operation of extraction and refolding process which is barely practical on large scale production.

Mammalian cells to produce tPA that resembles the endogenous protein found in circulation in small quantities. This is to mean that mammalian cell cultures are required in large amounts to produce the tPA required by the market demand.The utilization of recombinant protein therapeutics has transformed the aspect of modern pharmaceuticals. There is continued provision of innovative and effective therapies for a number of beforehand stubborn infections. The proteins are produced in large-scale through cultivation of genetically modified “host” cells, which harbour genes encoding for the desire proteins transfected artificially.

The effectiveness of the protein therapeutics is based on their production being in biologically active forms. This is achieved through proper folding and post-translational modifications. From the tPA bioprocessing, it is clear that it is not always expensive to utilize mammalian cell culture compared to prokaryotic cultures. A thorough economic analysis of the two processes should be done and more so using different pathways. The most effective pathway and cell culture should be used to make the mediation available in desired amounts and at an affordable price for all.

The proteins produced in mammalian cell cultures resemble those found in indigenous hence are bound to be more effective. This can be utilised in the present day biotechnology research for the production of other proteins. Research is to be directed on ways of improving the gram per litre range in order to improve on cost efficiency. ReferencesChan, V. L.,et al.(1972).Mammalian cells with altered forms of RNA polymerase II. Proc. Natl. Acad. Sci. USA: 69(11), pp. 3119 -123 Yerganian, G.(1985).

The biology and genetics of Chinese hamster.Molecular Cell Genetics. New York: John Wiley & Sons, Inc. pp. 3 - 36 Tjio, J. H. and T. T. Puck.(1958). Genetics of somatic mammalian cells II.J. Exp. Med. 108(2), pp.259 - 268