So far, pharmaceuticals used for the treatment ofdiseases have been based largely on the natural orsynthetic organic molecules produced by microbes orsynthesised by organic chemists. However, the post- genomic-era scenario projects a remarkable change in the therapeutics market, with the realisation that proteins have many potential therapeutic advantages for preventing and curing diseases and disorders. Proteins are the ultimate players of cellularfunction. Information stored in DNA directs the protein-synthesising machinery of the cell to produce specific proteins required for its structure, function and regulation. There are many proteins that are essential for good health that some people cannot produce because of genetic defects. The advent of sophisticated genomics and proteomics- based functional identification has unravelled large numbers of candidate proteins with a potential fortherapeutic intervention. Several market research groups have predicted that the growth of the biopharmaceutical market might reach as high as 15% annually and result in a critical shortage in protein-manufacturing capacity in the near future. Short peptide chains consisting of fewer than 30 amino acids can be synthesised chemically. Proteins larger than that are best produced by living cells. These ‘natural bioreactors’ support the sustained yield of target protein. The protein-encoding DNA is inserted into cells. As the cells grow, they synthesise the protein, which is subsequently harvested and purified. Production of proteins by micro-organisms is well entrenched in the industry. More recent knowledge of the molecular and cellular mechanism of diverse hosts has led to envisioning the art of expression offoreign proteins in mammalian cell culture and transgenic animals and plants. However, several key issues, namely complexity of the protein, quantity requirement and reliability of the expression system, need to be assessed before choosing a host system fora particular therapeutic protein. Microbial and plant hosts can be utilised for a moderate and large quantity of low-complexity proteins. Complex proteins can be produced in relatively small and large quantities in animal cell culture and transgenic animals, respectively. Since 1982, more than 100 therapeutic proteins and peptides have been licensed for production using bacterial, fungal and mammalian cells, and many therapeutic proteins are currently being developed and tested in a variety of host cells. 1 An overview ofthe technologies available and technical hurdles that are encountered in these processes is presented here. Production of Low-complexity Proteins Mi c r o b i a l C e l l F a c t o r i e s Humans have exploited microbial transformations for centuries – initially only in the food industry, but, lately, they have provided immense benefits to the pharmaceutical industry as bioreactors forantibiotic production. The birth of genetic engineering in the late 1970s, and subsequent technological expansion, has provided moleculartools for producing heterologous proteins in a wide range of micro-organisms including bacteria, yeast and filamentous fungi, thus largely extending theirmanufacturing capabilities. 2 Following transformation of a best protein- expressing host (for example Escherichia coli, Saccharomyces cerevisiaeand Pichia pastoris), best protein-expressing clones are identified and furtheroptimised for protein expression. Because ofscalability and well-characterised genetics, microbial host systems remain the best option for low- complexity, moderate-volume protein production at relatively low cost. 3 A major hurdle in microbial 1. R Andersson and R Mynahan, “The Prote in Production Ch allenge”, http:/ /www.windhov er.com 2. “Old bugs for new tasks; the microbial offer in the proteomics era”, http:// www.microbi alcellfactories .com/content/1/1 /4 3. R Andersson and R Manahan , “The Protein Production Challenge”, In Vivo, Windhover Information Inc., May 2001, pp. i–5. Dr Nilanjan Roy Shruti Agarwal Dr Nilanjan Roy is Head of the Proteomics Laboratory of the National Institute of Pharmaceutical Education and Research (NIPER). In addition to research, he is actively engaged in teaching in the areas of advanced techniques in biotechnology, genomics, proteomics, bio and pharma informatics. As well as his research articles, he is the author of several reports and popular articles. Dr Roy holds a PhD from Bose Institute, Kolkata, India, and has seven years of research experience from one of the premier hospitals of the US, The Cleveland Clinic Foundation, Cleveland, OH. Shruti Agarwal is an active researcher at NIPER’s Proteomics Laboratory. She has been involved in deciphering the role ofoxidative stress regulators in the ageing process. a report by Dr Nilanjan Roy andShruti Agarwal Head and Researcher, Proteomics Laboratory, National Institute of Pharmaceutical Education and Research (NIPER) Therapeutic Protein Production – An Overview BUSINESS BRIEFING: FUTURE DRUG DISCOVERY 2003 79 Technolo gy PROTEOMES & PROTEOMICS
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