Glacial lake outburst floods threaten millions globally.

Glacial lake outburst floods (GLOFs) represent a major hazard and can result in significant loss of life. Globally, since 1990, the number and size of glacial lakes has grown rapidly along with downstream population, while socio-economic vulnerability has decreased. Nevertheless, contemporary exposure and vulnerability to GLOFs at the global scale has never been quantified. Here we show that 15 million people globally are exposed to impacts from potential GLOFs. Populations in High Mountains Asia (HMA) are the most exposed and on average live closest to glacial lakes with ~1 million people living within 10 km of a glacial lake. More than half of the globally exposed population are found in just four countries: India, Pakistan, Peru, and China. While HMA has the highest potential for GLOF impacts, we highlight the Andes as a region of concern, with similar potential for GLOF impacts to HMA but comparatively few published research studies.

Assessing climate change associated sea-level rise impacts on sea turtle nesting beaches using drones, photogrammetry and a novel GPS system.

Climate change associated sea-level rise (SLR) is expected to have profound impacts on coastal areas, affecting many species, including sea turtles which depend on these habitats for egg incubation. Being able to accurately model beach topography using digital terrain models (DTMs) is therefore crucial to project SLR impacts and develop effective conservation strategies. Traditional survey methods are typically low-cost with low accuracy or high-cost with high accuracy. We present a novel combination of drone-based photogrammetry and a low-cost and portable real-time kinematic (RTK) GPS to create DTMs which are highly accurate (<10 cm error) and visually realistic. This methodology is ideal for surveying coastal sites, can be broadly applied to other species and habitats, and is a relevant tool in supporting the development of Specially Protected Areas. Here, we applied this method as a case-study to project three SLR scenarios (0.48, 0.63 and 1.20 m) and assess the future vulnerability and viability of a key nesting habitat for sympatric loggerhead (Caretta caretta) and green turtle (Chelonia mydas) at a key rookery in the Mediterranean. We combined the DTM with 5 years of nest survey data describing location and clutch depth, to identify (a) regions with highest nest densities, (b) nest elevation by species and beach, and (c) estimated proportion of nests inundated under each SLR scenario. On average, green turtles nested at higher elevations than loggerheads (1.8 m vs. 1.32 m, respectively). However, because green turtles dig deeper nests than loggerheads (0.76 m vs. 0.50 m, respectively), these were at similar risk of inundation. For a SLR of 1.2 m, we estimated a loss of 67.3% for loggerhead turtle nests and 59.1% for green turtle nests. Existing natural and artificial barriers may affect the ability of these nesting habitats to remain suitable for nesting through beach migration.

Leveraging single-pass tangential flow filtration to enable decoupling of upstream and downstream monoclonal antibody processing.

Decoupling upstream and downstream operations in biopharmaceutical production could enable more flexible manufacturing operations and could allow companies to leverage strategic or financial benefits that would be otherwise unattainable. A decoupling process was developed and scaled up utilizing single-pass tangential flow filtration for volume reduction, followed by bulk freezing in single-use bags prior to purification. Single-pass tangential flow filtration can be used to continuously concentrate harvested cell culture fluid, reducing the volume by 15-25× with a step yield of >96%. These concentration factors were reproduced with a second product, indicating that the process could be amenable to platform processes. Experimental data indicate that the product tested was stable for at least one year at -40 or -70°C. The concentration of the harvested cell culture fluid-either with or without a subsequent period of frozen storage-had no impact on the product quality attributes that were tested. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:405-411, 2018.

Reexamining opportunities for therapeutic protein production in eukaryotic microorganisms.

Antibodies are an important class of therapeutics and are predominantly produced in Chinese Hamster Ovary (CHO) cell lines. While this manufacturing platform is sufficiently productive to supply patient populations of currently approved therapies, it is unclear whether or not the current CHO platform can address two significant areas of need: affordable access to biologics for patients around the globe and production of unprecedented quantities needed for very large populations of patients. Novel approaches to recombinant protein production for therapeutic biologic products may be needed, and might be enabled by non-mammalian expression systems and recent advances in bioengineering. Eukaryotic microorganisms such as fungi, microalgae, and protozoa offer the potential to produce high-quality antibodies in large quantities. In this review, we lay out the current understanding of a wide range of species and evaluate based on theoretical considerations which are best poised to deliver a step change in cost of manufacturing and volumetric productivity within the next decade.Related article: http://onlinelibrary.wiley.com/doi/10.1002/bit.26383/full.

Mid-Holocene pulse of thinning in the Weddell Sea sector of the West Antarctic ice sheet.

Establishing the trajectory of thinning of the West Antarctic ice sheet (WAIS) since the last glacial maximum (LGM) is important for addressing questions concerning ice sheet (in)stability and changes in global sea level. Here we present detailed geomorphological and cosmogenic nuclide data from the southern Ellsworth Mountains in the heart of the Weddell Sea embayment that suggest the ice sheet, nourished by increased snowfall until the early Holocene, was close to its LGM thickness at 10 ka. A pulse of rapid thinning caused the ice elevation to fall ∼400 m to the present level at 6.5-3.5 ka, and could have contributed 1.4-2 m to global sea-level rise. These results imply that the Weddell Sea sector of the WAIS contributed little to late-glacial pulses in sea-level rise but was involved in mid-Holocene rises. The stepped decline is argued to reflect marine downdraw triggered by grounding line retreat into Hercules Inlet.

Evidence for the stability of the West Antarctic Ice Sheet divide for 1.4 million years.

Past fluctuations of the West Antarctic Ice Sheet (WAIS) are of fundamental interest because of the possibility of WAIS collapse in the future and a consequent rise in global sea level. However, the configuration and stability of the ice sheet during past interglacial periods remains uncertain. Here we present geomorphological evidence and multiple cosmogenic nuclide data from the southern Ellsworth Mountains to suggest that the divide of the WAIS has fluctuated only modestly in location and thickness for at least the last 1.4 million years. Fluctuations during glacial-interglacial cycles appear superimposed on a long-term trajectory of ice-surface lowering relative to the mountains. This implies that as a minimum, a regional ice sheet centred on the Ellsworth-Whitmore uplands may have survived Pleistocene warm periods. If so, it constrains the WAIS contribution to global sea level rise during interglacials to about 3.3 m above present.

Effects of solution environment on mammalian cell fermentation broth properties: enhanced impurity removal and clarification performance.

The processing of recombinant proteins from high cell density, high product titer cell cultures containing mammalian cells is commonly performed using tangential flow microfiltration (MF). However, the increased cellular debris present in these complex feed streams can prematurely foul the membrane, adversely impacting MF capacity and throughput. In addition, high cell density cell culture streams introduce elevated levels of process-related impurities, which increase the burden on subsequent purification operations to remove these complex media components and impurities. To address this challenge, an evaluation of mammalian cell culture broth buffer properties was examined to determine if enhanced impurity removal and clarification performance could be achieved. A framework is presented here for establishing optimized mammalian cell culture buffer conditions, involving trade-offs between product recovery and purification and improved clarification at manufacturing-scale production. A reduction in cell culture broth pH to 4.7-5.0 induced flocculation and impurity precipitation which increased the average feed particle-size. These conditions led to enhanced impurity removal and improved MF throughput and filter capacity for several mammalian systems. Feed conditions were further optimized by controlling ionic composition along with pH to improve product recovery from high cell density/high product titer cell cultures.

Modeling industrial centrifugation of mammalian cell culture using a capillary based scale-down system.

Continuous-flow centrifugation is widely utilized as the primary clarification step in the recovery of biopharmaceuticals from cell culture. However, it is a challenging operation to develop and characterize due to the lack of easy to use, small-scale, systems that can be used to model industrial processes. As a result, pilot-scale continuous centrifugation is typically employed to model large-scale systems requiring a significant amount of resources. In an effort to reduce resource requirements and create a system which is easy to construct and utilize, a capillary shear device, capable of producing energy dissipation rates equivalent to those present in the feed zones of industrial disk stack centrifuges, was developed and evaluated. When coupled to a bench-top, batch centrifuge, the capillary device reduced centrate turbidity prediction error from 37% to 4% compared to using a bench-top centrifuge alone. Laboratory-scale parameters that are analogous to those routinely varied during industrial-scale continuous centrifugation were identified and evaluated for their utility in emulating disk stack centrifuge performance. The resulting relationships enable bench-scale process modeling of continuous disk stack centrifuges using an easily constructed, scalable, capillary shear device coupled to a typical bench-top centrifuge.

Downstream antibody purification using aqueous two-phase extraction.

The extraction of antibodies using a polyethylene glycol (PEG)-citrate aqueous two-phase system (ATPS) was investigated. Studies using purified monoclonal antibody (mAb) identified operating ranges for successful phase formation and factors that significantly affected antibody partitioning. The separation of antibody and host cell protein (HCP) from clarified cell culture media was examined using statistical design of experiments (DOE). The partitioning of antibody was nearly complete over the entire range of the operating space examined. A model of the HCP partitioning was generated in which both NaCl and citrate concentrations were identified as significant factors. To achieve the highest purity, the partitioning of HCP from cell culture fluid into the product containing phase was minimized using a Steepest Descent algorithm. An optimal ATPS consisting of 14.0% (w/w) PEG, 8.4% (w/w) citrate, and 7.2% (w/w) NaCl at pH 7.2 resulted in a product yield of 89%, an approximate 7.6-fold reduction in HCP levels relative to the clarified cell culture fluid before extraction and an overall purity of 70%. A system consisting of 15% (w/w) PEG, 8% (w/w) citrate, and 15% (w/w) NaCl at pH 5.5 reduced product-related impurities (aggregates and low molecular product fragments) from ∼40% to less than 0.5% while achieving 95% product recovery. At the experimental conditions that were optimized in the batch mode, a scale-up model for the use of counter-current extraction technology was developed to identify potential improvements in purity and recovery that could be realized in the continuous operational mode.

Concentration of proteins and removal of solutes.

The dramatic advances in recombinant DNA and proteomics technology over the past decade have supported a tremendous growth in biologics applied to diagnostics, biomarkers, and commercial therapeutic markets. In particular, antibodies and fusion proteins have now become a main focus for a broad number of clinical indications, including neurology, oncology, and infectious diseases with projected increase in novel first-class molecules and biosimilar entities over the next several years. In line with these advances are the improved analytical, development, and small-scale preparative methods employed to elucidate biologic structure, function, and interaction. A number of established methods are used for solvent removal, including lyophilization, reverse extraction, solute precipitation, and dialysis (solvent exchange), ultrafiltration, and chromatographic techniques. Notably, advances in the miniaturization and throughput of protein analysis have been supported by the development of a plethora of microscale extraction procedures and devices that exploit a wide array of modes for small-scale sample preparation, including the concentration and desalting of protein samples prior to further analysis. Furthermore, advances in process handling and data monitoring at microscale have dramatically improved complex control and product recovery of small quantities of biologics using techniques such as lyophilization and precipitation. In contrast, the efficient concentration of feed streams during preparative chromatography has been enhanced by improvements to protein binding capacity achieved through advanced bead and ligand design. The objective of solvent removal may be to prepare or concentrate solutes for analysis, or to facilitate their production or modification. Here, we describe the most recent advances in these techniques, particularly focusing on improved capabilities for bench-scale preparative methods.