Cell-Free Expression for Nanolipoprotein Particles: Building a High-Throughput Membrane Protein Solubility Platform

Cappuccio, Jenny A.; Hinz, Angela K.; Kuhn, Edward A.; Fletcher, Julia E.; Arroyo, Erin S.; Henderson, Paul; Blanchette, Craig D.; Walsworth, Vickie L.; Corzett, Michele; Law, Richard; Pesavento, Joseph B.; Segelke, Brent W.; Sulchek, Todd; Chromy, Brett A.; Katzen, Federico; Peterson, Todd C.; Bench, Graham; Kudlicki, Wiesław; Hoeprich, Paul D.; Coleman, Matthew A.

doi:10.1007/978-1-59745-196-3_18

Cited by 39 publications

(30 citation statements)

References 46 publications

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“…Using this approach, two membrane proteins, the tetracycline pump (TetA) and mannitol permease (MtlA), were expressed and achieved the high yield of 570 and 130 μg mL -1 respectively, up to 400 times as previous methods (Wuu and Swartz, 2008). Nanolipoprotein particles, which are lipid bilayers confined within a ring of amphipathic protein of defined diameter (Cappuccio et al , 2009), as well as unilamellar liposomes (Goren et al , 2009) have also shown tremendous promise.…”

Section: Applicationsmentioning

confidence: 99%

Cell-free protein synthesis: Applications come of age

Carlson

Gan

Hodgman

et al. 2012

Biotechnology Advances

594

559

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Cell-free protein synthesis has emerged as a powerful technology platform to help satisfy the growing demand for simple and efficient protein production. While used for decades as a foundational research tool for understanding transcription and translation, recent advances have made possible cost-effective microscale to manufacturing scale synthesis of complex proteins. Protein yields exceed grams protein produced per liter reaction volume, batch reactions last for multiple hours, costs have been reduced orders of magnitude, and reaction scale has reached the 100-liter milestone. These advances have inspired new applications in the synthesis of protein libraries for functional genomics and structural biology, the production of personalized medicines, and the expression of virus-like particles, among others. In the coming years, cell-free protein synthesis promises new industrial processes where short protein production timelines are crucial as well as innovative approaches to a wide range of applications.

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Section: Applicationsmentioning

confidence: 99%

Cell-free protein synthesis: Applications come of age

Carlson

Gan

Hodgman

et al. 2012

Biotechnology Advances

594

559

View full text Add to dashboard Cite

show abstract

“…This method allows for engineering of lipoprotein composition at the molecular level, opening the door for any number of membrane-compatible components to be included in the nanoparticle. Proton pumps [120, 121], membrane-associating enzymes [122], and signaling proteins [118, 123] have all been successfully incorporated into lipoprotein nanostructure.…”

Section: Cell-membrane Mimicking Nanoparticles For Toxin Neutralizmentioning

confidence: 99%

Engineered nanoparticles mimicking cell membranes for toxin neutralization

Fang

Luk

et al. 2015

Advanced Drug Delivery Reviews

112

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Protein toxins secreted from pathogenic bacteria and venomous animals rely on multiple mechanisms to overcome the cell membrane barrier to inflict their virulence effect. A promising therapeutic concept toward developing a broadly applicable anti-toxin platform is to administer cell membrane mimics as decoys to sequester these virulence factors. As such, lipid membrane-based nanoparticulates are an ideal candidate given their structural similarity to cellular membranes. This article reviews the virulence mechanisms employed by toxins at the cell membrane interface and highlights the application of cell-membrane mimicking nanoparticles as toxin decoys for systemic detoxification. In addition, the implication of particle/toxin nanocomplexes in the development of toxoid vaccines is discussed.

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“…The structure and function of HDLs in vivo have been studied for the past three decades, and methods for assembling several different compositionally distinct HDLs ex vivo [also called reconstituted HDLs (rHDLs), nanodiscs, or nanolipoprotein particles (NLPs)] have been developed [11]–[14]. The vast majority of the work on rHDLs and NLPs has been directed at both understanding the biology of such particles [15]–[18] as well as exploring their utility in solubilizing and stabilizing membrane proteins in discrete, native lipid environments [19]–[24]. However, the use of these particles for delivery of therapeutic drugs [25]–[28], diagnostic imaging [29], and vaccine and immunomodulation applications [30]–[33] has only recently been examined.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Nanolipoprotein Particles (NLPs) as an In Vivo Delivery Platform

et al. 2014

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Nanoparticles hold great promise for the delivery of therapeutics, yet limitations remain with regards to the use of these nanosystems for efficient long-lasting targeted delivery of therapeutics, including imparting functionality to the platform, in vivo stability, drug entrapment efficiency and toxicity. To begin to address these limitations, we evaluated the functionality, stability, cytotoxicity, toxicity, immunogenicity and in vivo biodistribution of nanolipoprotein particles (NLPs), which are mimetics of naturally occurring high-density lipoproteins (HDLs). We found that a wide range of molecules could be reliably conjugated to the NLP, including proteins, single-stranded DNA, and small molecules. The NLP was also found to be relatively stable in complex biological fluids and displayed no cytotoxicity in vitro at doses as high as 320 µg/ml. In addition, we observed that in vivo administration of the NLP daily for 14 consecutive days did not induce significant weight loss or result in lesions on excised organs. Furthermore, the NLPs did not display overt immunogenicity with respect to antibody generation. Finally, the biodistribution of the NLP in vivo was found to be highly dependent on the route of administration, where intranasal administration resulted in prolonged retention in the lung tissue. Although only a select number of NLP compositions were evaluated, the findings of this study suggest that the NLP platform holds promise for use as both a targeted and non-targeted in vivo delivery vehicle for a range of therapeutics.

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Cell-Free Expression for Nanolipoprotein Particles: Building a High-Throughput Membrane Protein Solubility Platform

Cited by 39 publications

References 46 publications

Cell-free protein synthesis: Applications come of age

Cell-free protein synthesis: Applications come of age

Engineered nanoparticles mimicking cell membranes for toxin neutralization

Evaluation of Nanolipoprotein Particles (NLPs) as an In Vivo Delivery Platform

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