Recombinant proteins large-scale production

R)- and ( -selective HNLs. A number of recombinant HNLs have also been expressed in E. coli, Saccharomyces cerevisiae, and Pichia pastoris. Recently, protein engineering has been successfully applied to the development of a tailor-made HNL for large-scale production of specific cyanohydrins [69,70]. 

Lundstrom, K. (2003) Semliki Forest vims vectors for large-scale production of recombinant proteins. Methods... 

Large-Scale Production of Proteins from Recombinant DNA Using Suspensions of Animal Cells... 

Despite these theoretical concerns, a number of HIV vaccines are under development. Most of these vaccines have been developed by recombinant DNA techniques that have allowed a large-scale production of individual viral proteins. The predominant HIV proteins that make up these potential vaccines are env proteins (e.g., gp 120) and, to a lesser extent, gag proteins. In addition, inactivated whole HIV virus is being tested. 

Mammalian cells are commonly employed for the production of therapeutic and diagnostic proteins, since they are able to correctly synthetize the large and complex structures that the human body requires as medicine [1]. Nowadays, they are employed for the large-scale production of recombinant therapeutic proteins, monoclonal antibodies (MAbs) and viruses used in the preparation of vaccines (e.g. against rabies, hepathytis B, polio, etc) [2]. An overview of some licensed/approved products derived from mammalian cell culture is given in Table 1. 

The advent of recombinant DNA technology has allowed the isolation of genes and expression of proteins that are found in biologic tissues in exceedingly small quantities. This has permitted large-scale production of enzymes such as tissue plasminogen activator (i.e., tPA, alteplase) that cannot be extracted from tissues in quantities required for therapeutic use. Table 9.2... 

Although numerous cell lines have been screened for their efficiency as a host system for recombinant protein production, only a few have shown favorable properties for the expression of biopharmaceuticals (Hauser, 1997). Regulatory and economic issues for large-scale production and the intended application of the recombinant protein (diagnosis, therapy, etc.) have to be carefully considered (Makrides and Prentice, 2003). Three mammalian cell lines are now commonly used by the pharmaceutical industry Chinese hamster ovary (CHO) cells, the murine myeloma SP2/0 and the NS0 cell line (see Table 3.1). These cell lines have been used to produce 11 of 21 therapeutic products approved from 1996 to 2000 (Chu and Robinson, 2001). 

We prefer large-scale production of recombinant protein by infection in mono-layer culture, since it is easier to achieve synchronous infections in monolayer culture than in suspension culture. 

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