Analysis of fouling and breakthrough of process related impurities during depth filtration using confocal microscopy

Titer improvement has driven process intensification in mAb manufacture. However, this has come with the drawback of high cell densities and associated process related impurities such as cell debris, host cell protein (HCP), and DNA. This affects the capacity of depth filters and can lead to carryover of impurities to protein A chromatography leading to early resin fouling. New depth filter materials provide the opportunity to remove more process related impurities at this early stage in the process. Hence, there is a need to understand the mechanism of impurity removal within these filters. In this work, the secondary depth filter Millistak+ X0HC (cellulose and diatomaceous earth) is compared with the X0SP (synthetic), by examining the breakthrough of DNA and HCP. Additionally, a novel method was developed to image the location of key impurities within the depth filter structure under a confocal microscope. Flux, tested at 75, 100, and 250 LMH was found to affect the maximal throughput based on the max pressure of 30 psi, but no significant changes were seen in the HCP and DNA breakthrough. However, a drop in cell culture viability, from 87% to 37%, lead to the DNA breakthrough at 10% decreasing from 81 to 55 L/m2 for X0HC and from 105 to 47 L/m2 for X0SP. The HCP breakthrough was not affected by cell culture viability or filter type. The X0SP filter has a 30%–50% higher max throughput depending on viability, which can be explained by the confocal imaging where the debris and DNA are distributed differently in the layers of the filter pods, with more of the second tighter layer being utilized in the X0SP

Hybrid Flooding Scheme for Mobile Wireless Networks

A hybrid broadcast scheme for mobile wireless networks is proposed in this letter. The main objective is to combine different flooding schemes in order to solve the broadcast storm issue encountered by the simple flooding scheme. For this purpose, the density of nodes is taken into account using a density metric called expansion metric. In addition, in order to reduce the broken links due to mobility of nodes and increasing dissimilarity among the intermediate nodes, a forwarding zone criterion is included in the proposed schemes. The proposed approaches have been implemented and compared with pure probabilistic flooding, and simple flooding schemes

High-resolution imaging of depth filter structures using X-ray computed tomography

A multiple length scale approach to the imaging and measurement of depth filters using X-ray computed tomography is described. Three different filter grades of varying nominal retention ratings were visualized in 3D and compared quantitatively based on porosity, pore size and tortuosity. Positional based analysis within the filters revealed greater voidage and average pore sizes in the upstream quartile before reducing progressively through the filter from the center to the downstream quartile, with these results visually supported by voidage distance maps in each case. Flow simulation to display tortuous paths that flow may take through internal voidage were examined. Digital reconstructions were capable of identifying individual constituents of voidage, cellulose and perlite inside each depth filter grade, with elemental analysis on upstream and downstream surfaces confirming perlite presence. Achieving an appropriate pixel size was of particular importance when optimizing imaging conditions for all grades examined. A 3 µm pixel size was capable of representing internal macropores of each filter structure; however, for the finest grade, an improvement to a 1 µm pixel size was required in order to resolve micropores and small perlite shards. Enhancing the pixel size resulted in average porosity measurements of 70% to 80% for all grades. Graphical abstract: [Figure not available: see fulltext.

3DFeat-Net: Weakly Supervised Local 3D Features for Point Cloud Registration

In this paper, we propose the 3DFeat-Net which learns both 3D feature detector and descriptor for point cloud matching using weak supervision. Unlike many existing works, we do not require manual annotation of matching point clusters. Instead, we leverage on alignment and attention mechanisms to learn feature correspondences from GPS/INS tagged 3D point clouds without explicitly specifying them. We create training and benchmark outdoor Lidar datasets, and experiments show that 3DFeat-Net obtains state-of-the-art performance on these gravity-aligned datasets.Comment: 17 pages, 6 figures. Accepted in ECCV 201

Topological descriptors for 3D surface analysis

We investigate topological descriptors for 3D surface analysis, i.e. the classification of surfaces according to their geometric fine structure. On a dataset of high-resolution 3D surface reconstructions we compute persistence diagrams for a 2D cubical filtration. In the next step we investigate different topological descriptors and measure their ability to discriminate structurally different 3D surface patches. We evaluate their sensitivity to different parameters and compare the performance of the resulting topological descriptors to alternative (non-topological) descriptors. We present a comprehensive evaluation that shows that topological descriptors are (i) robust, (ii) yield state-of-the-art performance for the task of 3D surface analysis and (iii) improve classification performance when combined with non-topological descriptors.Comment: 12 pages, 3 figures, CTIC 201

Learning and Matching Multi-View Descriptors for Registration of Point Clouds

Critical to the registration of point clouds is the establishment of a set of accurate correspondences between points in 3D space. The correspondence problem is generally addressed by the design of discriminative 3D local descriptors on the one hand, and the development of robust matching strategies on the other hand. In this work, we first propose a multi-view local descriptor, which is learned from the images of multiple views, for the description of 3D keypoints. Then, we develop a robust matching approach, aiming at rejecting outlier matches based on the efficient inference via belief propagation on the defined graphical model. We have demonstrated the boost of our approaches to registration on the public scanning and multi-view stereo datasets. The superior performance has been verified by the intensive comparisons against a variety of descriptors and matching methods

Analytical tools for monitoring changes in physical and chemical properties of chromatography resin upon reuse

Protein A resins are often reused for multiple cycles to improve process economy during mAb purification. Significant reduction in binding capacity and product recovery are typically observed due to the presence of unwanted materials (foulants) deposited on the resin upon reuse. In this paper, we have used a wide spectrum of qualitative and quantitative analytical tools (particle size analysis, HPLC, fluorescence, SEM, MS, and FTIR) to compare the strengths and shortcomings of different analytical tools in terms of their capability to detect the fouling of the resin and relate it to chromatographic cycle performance. While each tool offers an insight into this complex phenomena, fluorescence is the only one that can be used for real‐time monitoring of resin fouling. A correlation could be established between fluorescence intensity and the process performance attributes (like yield or binding capacity) impacted upon resin reuse. This demonstration of the application of fluorescence for real‐time monitoring correlated empirically with process performance attributes and the results support its use as a PAT tool as part of a process control strategy. While the focus of this paper is on fouling of protein A chromatography resin, the approach and strategy are pertinent to other modes of chromatography as well

Packed bed compression visualisation and flow simulation using an erosion-dilation approach

X-ray computed tomography has been demonstrated to be capable of imaging 1 mL (5 mm diameter, 50 mm height) chromatography packed beds under compression, visualising the 3D structure and measuring changes to geometry of the packing. 1 mL pre-packed columns did not exhibit any structural changes at vendor specified flow rate limits, however cellulose beds did compress at higher flow rates that were imaged before, during and after flow. This was used to visualise and quantitate changes to porosity, tortuosity and permeability based on simulation of flow through the packed bed structure using the imaging data. When using a high flow rate it was found that a decrease in porosity could be measured during compression before reverting after flow had ceased, with corresponding changes to tortuosity and permeability also occurring. X-ray CT imaging of packed beds and individual beads exposed to foulant-rich process streams resulted in considerable image quality loss, associated with residual biological material. In order to address this, digital processing using an erosion-dilation method was applied at bead and bed scales to computationally alter the porosity by adding or removing material from the existing surface to calculate the impact upon tortuosity factor. The eroded and dilated bead volumes of agarose, cellulose and ceramic materials were used to simulate diffusivity whilst mimicking internal bead pore constriction and blocking mechanisms

Experiments in no-impact control of dingoes: Comment on Allen et al. 2013

There has been much recent debate in Australia over whether lethal control of dingoes incurs environmental costs, particularly by allowing increase of populations of mesopredators such as red foxes and feral cats. Allen et al. (2013) claim to show in their recent study that suppression of dingo activity by poison baiting does not lead to mesopredator release, because mesopredators are also suppressed by poisoning. We show that this claim is not supported by the data and analysis reported in Allen et al.'s paper. © 2014 Johnson et al.; licensee BioMed Central Ltd