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Manufacture of the Human Growth Hormone

359 words | 2 page(s)

The production of the human growth hormone (hGH) requires the use of recombinant DNA (rDNA) technology (Brennan, 2017). Facilities required include a laboratory for production of rDNA. A plasmid vector and E. coli are required for the creation of rDNA. Fermentation chambers are required mass cultivation of E. coli and filtration chamber are required for separation and purification of the hGH. The process involves the isolation of genes, specifically DNA which codes for hGH. These pieces are then attached to other pieces of DNA, and the resultant product is transferred to another species. The species that is mostly used in this case is the bacteria with Escherichia coli (E. coli) being the most popular host for the production (Rezaei & Zarkesh-Esfahani, 2012).

Once the genes are inserted into bacteria, plasmid, which acts as a vector with the information required for the complete cycle, is then introduced. The next stages involve the fermentation, harvesting, followed by the purification of the hormone for use in the intended purpose. Fermentation comprises of three major steps: inoculation, seed fermentation, and production fermentation (Kanner & Schmell, 2010). Inoculation involves the insertion of the host bacteria into a flask where it remains for six hours. The flask contains growth elements whose quantity is determined by the quantity of the hGH anticipated. NaCl, ampicillin sodium salt, casein hydrolysate and, yeast extract. Seed fermentation involves continuous fermentation at temperatures of 300C and PH levels of 7. The same growth elements used in the inoculation stage are used, and some other elements added, including glucose and antifoam (Kanner & Schmell, 2010). Production fermentation involves the maintenance of all elements utilized in seed fermentation and additional 50% of glucose solution.

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The temperature and PH levels of the seed fermentation stage are also maintained. The temperature is then raised to 420C and fermentation continued for another two hours (Kanner & Schmell, 2010). The hGH is then harvested using the hollow fiber microfiltration method. The recovery of the hGH involves the disruption of the harvest cell slurry and washing with purified water to obtain inclusion bodies. The latter is then dissolved in NaOH and the PH raised to 12 while stirring. The hGH can then be purified using chromatography or ultrafiltration (Kanner & Schmell, 2010).

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