2016
DOI: 10.1080/21655979.2016.1241925
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The growing impact of lyophilized cell-free protein expression systems

Abstract: Recently reported shelf-stable, on-demand protein synthesis platforms are enabling new possibilities in biotherapeutics, biosensing, biocatalysis, and high throughput protein expression. Lyophilized cell-free protein expression systems not only overcome cold-storage limitations, but also enable stockpiling for ondemand synthesis and completely sterilize the protein synthesis platform. Recently reported high-yield synthesis of cytotoxic protein Onconase from lyophilized E. coli extract preparations demonstrates… Show more

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Cited by 43 publications
(31 citation statements)
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“…There has recently been a surge of interest in ‘in vitro biology’ [ 38 40 ], with in vitro transcription and translation (IVTT) reactions being used for diagnostic applications [ 41 ] and for the production of many new proteins [ 42 45 ]. Simplification of the traditionally labor and cost intensive procedures for preparation and storage of E .…”
Section: Discussionmentioning
confidence: 99%
“…There has recently been a surge of interest in ‘in vitro biology’ [ 38 40 ], with in vitro transcription and translation (IVTT) reactions being used for diagnostic applications [ 41 ] and for the production of many new proteins [ 42 45 ]. Simplification of the traditionally labor and cost intensive procedures for preparation and storage of E .…”
Section: Discussionmentioning
confidence: 99%
“…The activity of cell-free-derived sfGFP was determined using an in-extract fluorescence analysis as described previously (39). Briefly, 2 μL of cell-free reaction product was diluted into 48 μL of Ambion nanopure water (Invitrogen, USA).…”
Section: Gfp Fluorescence Assaymentioning
confidence: 99%
“…A body of work dedicated to optimization of extract preparation and reaction conditions has simplified, expedited, and improved the cost and performance of E. coli CFE systems (22,28,29). Optimized E. coli-based CFE reactions: (i) quickly synthesize grams of protein per liter in batch reactions (30)(31)(32), (ii) are scalable from the nL to 100 L scale (33,34), and (iii) can be freeze-dried for months of shelf-stability and distribution to the point of care (6,22,29,(35)(36)(37)(38)(39)(40). Freeze-dried CFE systems are poised to make disruptive impacts in biotechnology, having already been leveraged for point-of-use biosensing (41)(42)(43)(44)(45)(46), therapeutic and vaccine production (37,38,47), and educational kits (22,(48)(49)(50).…”
Section: Main Text Introductionmentioning
confidence: 99%
“…Cell‐free protein synthesis (CFPS) offers an alternative platform for ASNase activity assessment which presents multiple advantages over traditional approaches. First, CFPS reactions can be rendered shelf stable, thus mitigating the need for cold‐chain storage of assay components, enabling high‐density storage and facile use . Second, the CFPS platform provides an open reaction environment which may be dynamically optimized and is reportedly robust against samples containing human blood .…”
Section: Introductionmentioning
confidence: 99%