Imperial/Cell-Free/Comparison
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Cell-Free: A Comparison of In Vivo and In Vitro Systems
| Non-infectious because of non-proliferative nature | Some strains may be pathogenic |
| Process is quick and simple requiring only preparation of cell extract and feeding solution and subsequent addition of DNA template | Process is laborious involving DNA cloning and transformation and protein expression |
| Good control can be achieved easily using modified reaction conditions such as addition of accessory elements or inhibitory factors | Less controllability because of the presence of endogenous substances and because cells do not survive extreme conditions |
| Both plasmid and linear DNAs and can be used as templates for expression | Plasmid DNAs are usually used. Linear DNAs are easily degraded by endogenous nucleases |
| Protein degradation is minimized by adding protease inhibitors | Synthesized proteins may be degraded by endogenous proteases |
| Toxic proteins can be synthesized in large quantities | Synthesis of toxic proteins may kill the cells |
| Proteins containing unnatural amino acids can be achieved | Difficult to produce proteins containing unnatural amino acids |
| Shorter lifespan since system cannot replicate | Longer lifespan since system can replicate |
| More expensive because of the constant need for nutrient and energy supply | Less expensive because of the ability of the system to generate energy from relatively cheap nutrient source |
| Less characterized, less experience of use in the laboratories | Better characterized, more experience of use in the laboratories |
