Innovation Management in the Postal Sector: What Are the Internal Factors Influencing Postal Operator’s Innovation Management?

Since the past decades, the postal sector is undergoing profound reform (liberalization). The major motivation for such reform processes was and still is the belief that liberalization and deregulation stimulates, among others, product and process innovations, both of which are important vehicles for productivity and economic growth. Against the background of such reform processes, postal operators need to increase their performance through innovation in order to gain competitive advantage and to ensure profitability and growth. Therefore, postal companies have to adapt their innovation management to the changing environment. Until today, there is little research on the adaption of postal operator’s innovation management. In this paper, we aim to describe the internal factors that influence postal operators’ innovation management. The goals of this article are 1) to illustrate the internal factors that are influencing an organization’s ability to manage innovation and 2) to analyze these factors for the particular setting of the postal industry

Ultrahigh-Throughput Screening of an Artificial Metalloenzyme using Double Emulsions

The potential for ultrahigh-throughput compartmentalization renders droplet microfluidics an attractive tool for the directed evolution of enzymes. Importantly, it ensures maintenance of the phenotype-genotype linkage throughout optimization, enabling reliable identification of improved mutants. The full potential of droplet microfluidics remains unexplored, however, as droplet sorting often relies on low-throughput, custom-made devices that typically only allow simultaneous analysis of two parameters. Here, we report an approach for ultrahigh-throughput screening of an artificial metalloenzyme in double emulsion droplets (DEs) using commercially-available fluorescence-activated cell sorters (FACS). This protocol was validated by screening a 400 double-mutant streptavidin library for ruthenium-catalyzed deallylation of allocprotected aminocoumarin. The most active variants, identified by next generation sequencing, were in good agreement with hits obtained using a 96-well plate procedure. These findings pave the way for the systematic implementation of FACS for the directed evolution of enzymes and will significantly expand the accessibility of ultrahighthroughput DE screening protocols

Ultrahigh‐Throughput Screening of an Artificial Metalloenzyme using Double Emulsions

The potential for ultrahigh-throughput compartmentalization renders droplet microfluidics an attractive tool for the directed evolution of enzymes. Importantly, it ensures maintenance of the phenotype-genotype linkage, enabling reliable identification of improved mutants. Herein, we report an approach for ultrahigh-throughput screening of an artificial metalloenzyme in double emulsion droplets (DEs) using commercially available fluorescence-activated cell sorters (FACS). This protocol was validated by screening a 400 double-mutant streptavidin library for ruthenium-catalyzed deallylation of an alloc-protected aminocoumarin. The most active variants, identified by next-generation sequencing, were in good agreement with hits obtained using a 96-well plate procedure. These findings pave the way for the systematic implementation of FACS for the directed evolution of (artificial) enzymes and will significantly expand the accessibility of ultrahigh-throughput DE screening protocols

Microfluidic size-based separation, enrichment and analysis of vesicles

Extracellular vesicles are considered as valuable biomarkers for various diseases. The isolation from their surrounding matrix, e.g. blood, is a crucial step for a detailed analysis of their size or constituents. Therefore, we present a microfluidic platform capable of size-based separation of vesicles with a wide dynamic range via deterministic lateral displacement and the direct on-chip analysis of sorted vesicles. Polydisperse samples of giant and large unilamellar vesicles as biological sample models were separated and enriched in designated regions in the channel. Total internal reflection fluorescence spectroscopy allowed for a discrimination and enumeration of nanovesicles labeled with different membrane markers

Droplet Microfluidics and Directed Evolution of Enzymes: an Intertwined Journey

Evolution is essential to the appearance of complexity and ultimately Life. It relies on the propagation of the properties, traits and characteristics that allow an organism to survive in a challenging environment. It is evolution that shaped our world over about four billion years by slow and iterative adaptation. While natural evolution based on selection is slow and gradual, directed evolution allows the fast and streamlined optimization of a phenotype under selective conditions. The potential of directed evolution for the discovery and optimization of enzymes is mostly limited by the throughput of the tools and methods available for screening. Over the past twenty years, versatile tools based on droplet microfluidics have been developed to address the need for higher throughput. In this review, we provide a chronological overview of the intertwined development of microfluidics droplet-based compartmentalization methods and in vivo directed evolution of enzymes

Droplet Microfluidics and Directed Evolution of Enzymes: An Intertwined Journey

Evolution is essential to the generation of complexity and ultimately life. It relies on the propagation of the properties, traits, and characteristics that allow an organism to survive in a challenging environment. It is evolution that shaped our world over about four billion years by slow and iterative adaptation. While natural evolution based on selection is slow and gradual, directed evolution allows the fast and streamlined optimization of a phenotype under selective conditions. The potential of directed evolution for the discovery and optimization of enzymes is mostly limited by the throughput of the tools and methods available for screening. Over the past twenty years, versatile tools based on droplet microfluidics have been developed to address the need for higher throughput. In this Review, we provide a chronological overview of the intertwined development of microfluidics droplet-based compartmentalization methods and in vivo directed evolution of enzymes.ISSN:1433-7851ISSN:1521-3773ISSN:0570-083

Synchronized Reagent Delivery in Double Emulsions for Triggering Chemical Reactions and Gene Expression

Microfluidic methods to form single emulsion and double emulsion (DE) droplets have greatly enhanced the toolbox for high throughput screening for cell or enzyme engineering and drug discovery. However, remaining challenges in the supply of reagents into these enclosed nanoliter compartments limit the applicability of droplet microfluidics. Here, we introduce a strategy for on-demand delivery of reactants in DEs. We use lipid vesicles as transport carriers, which are co-encapsulated in double emulsions and release their cargo upon addition of an external trigger, here the anionic surfactant SDS. The reagent present inside the lipid vesicles stays isolated from the remaining content of the DE vessel until SDS enters the DE lumen and solubilizes the lipid bilayer. We demonstrate the versatility of the method with two critical applications, chosen as representative assays for high throughput screening. First, we trigger enzymatic reactions after releasing a reactant and second, we encapsulate bacteria and induce gene expression at a delayed time. The presented technique is compatible with the high throughput analysis of individual DE droplets using conventional flow cytometry as well as with microfluidic time-resolved studies. The possibility of delaying and controlling reagent delivery in current high throughput compartmentalization systems will significantly extend their range of applications e.g. for directed evolution, and further improve their compatibility with biological systems

A multiplexed cell-free assay in double emulsion droplets

We introduce a novel on-chip assay, performed in water-in-oil-in-water double emulsions, that uses cell-free protein synthesis and artificial liposomes for the screening of membrane disrupting toxin DNA libraries according to their antimicrobial activity and host membrane safety. Our method represents a unique approach to the selection and potentially directed evolution of membrane-disrupting toxins

Synchronized Reagent Delivery in Double Emulsions for Triggering Chemical Reactions and Gene Expression

Microfluidic methods for the formation of single and double emulsion (DE) droplets allow for the encapsulation and isolation of reactants inside nanoliter compartments. Such methods have greatly enhanced the toolbox for high-throughput screening for cell or enzyme engineering and drug discovery. However, remaining challenges in the supply of reagents into these enclosed compartments limit the applicability of droplet microfluidics. Here, a strategy is introduced for on-demand delivery of reactants in DEs. Lipid vesicles are used as reactant carriers, which are co-encapsulated in double emulsions and release their cargo upon addition of an external trigger, here the anionic surfactant sodium dodecyl sulfate (SDS). The reagent present inside the lipid vesicles stays isolated from the remaining content of the DE vessel until SDS enters the DE lumen and solubilizes the vesicles’ lipid bilayer. The versatility of the method is demonstrated with two critical applications chosen as representative assays for high-throughput screening: the induction of gene expression in bacteria and the initiation of an enzymatic reaction. This method not only allows for the release of the lipid vesicle content inside DEs to be synchronized for all DEs but also for the release to be triggered at any desired time.ISSN:2366-960