Commercial-scale Manufacturing Research Articles

Microparticles for Suspension Culture of Mammalian Cells

The focus of this work is to develop a technology for the synthesis of polymer microcarriers that demonstrate mammalian cell culture adhesion on the surface of the microcarriers. Most mammalian cells are adherent in nature that requires multilayer vessels, large volume, expensive cell culture media, high manufacturing time, and high costs of cell culture supplies for the commercial-scale manufacturing of cells. The development of an efficient, scalable technology for producing large volumes of cells is a need in bioprocess industries to improve product potency. We developed a method of synthesizing soft biocompatible US FDA approved polymer based microparticle carrier system of approximately 260 ± 27 μm in diameter that serves as an adherent platform for human umbilical vein endothelial cells (HUVEC) to grow in suspension. Our preliminary experimental results showed that using the polymeric microcarrier system cell adhesion to the surface of the microcarriers was 2-3-fold higher than conventional cell culture flasks while using 10-fold lower cell culture media in a bioreactor than a tissue-culture treated flask. The survival of HUVEC on microparticles was confirmed by live cell staining (green fluorescent calcein AM), dead cell staining (ethidium homodimer-1), nuclear DAPI staining, actin cytoskeleton staining, confocal microscopy, and flow cytometry analysis. This technology will provide high cell culture productivity while reducing the costs of growing adherent cells.

Impact of Manufacturing Tolerance on Lithium Ion Electrode and Cell Physical Properties

In both laboratory and commercial scale manufacturing of lithium ion cells, variability is introduced in numerous stages of the production process. The acceptable tolerances on these produced parts can have a large impact on the final product’s reliability, repeatability, and functionality. In this study, the range of electrode porosity, electrode internal void volume, cell capacity, and capacity ratio that result from electrode coating and calendering tolerance are investigated and offered as one possible explanation for cell-to-cell and lot-to-lot performance variation. Based on an electrode coating tolerance of ± 0.4 mg/cm2 and calender tolerance of ± 3.0 μm the resulting range of physical properties is investigated. For a target positive electrode porosity of 30%, the resulting porosity can range from 19.6-38.6%. To account for this variation during the manufacturing process, as much as 41% excess or as little as 59% of the target electrolyte quantity should be added to cells to match the positive electrode void volume. Similar results are observed for a negative electrode of 40% target porosity, where a range from 30.8% to 48.0% porosity is possible. For the negative electrode as little as 72% up to 28% excess electrolyte should be added to fill the internal void space. Although the results are specific to each electrode composition, density, chemistry, and loading the presented process highlight possible variability of produced parts. These results are further magnified as cell designs moves toward higher power applications with thinner electrode coatings.

Evaluating automated buffer exchange protocols using Rotea™ counterflow centrifuge

Background & Aim Buffer exchange is an essential process in cell therapy manufacturing where the introduction of automation can reduce costs and enable commercial scale manufacture. Rotea™ (Scinogy), is a benchtop counterflow centrifuge specifically designed for cell therapy processing, that allows closed automated buffer exchange for small to medium size batches. To determine the capability of Rotea™, we compared the automated protocols to manual centrifugation for buffer exchange using cultured Jurkat cells and mesenchymal stromal cells (MSCs). Methods, Results & Conclusion Methods The details of starting material and wash buffer for various cell types are listed in Table 1. In both automated protocols, wash buffer and cells were loaded into the cell transfer bags and connected onto the single-use processing kit. The automated process starts with loading the cells into the cell chamber, where they were then washed with 40 ml of the buffer before being concentrated to a final volume of 10 ml. The automated processes were compared to two cycles of centrifugation and resuspension. The recovered cells were also plated for MTS assays to measure cell proliferation over 48 hours. Results Automated buffer exchange has a shorter processing time compared to the manual process. Cell viabilities were generally improved after buffer exchange and cell recoveries were close to 100% (Table 1). Cell proliferation rates were similar between manual and automated processes. Conclusions The closed and automated counterflow centrifuge is ideally suited to perform buffer exchange for various cell types with high cell recoveries and shorter processing times.

Immunoglobulin Gamma-Like Therapeutic Bispecific Antibody Formats for Tumor Therapy.

Bispecific antibodies (BsAbs) are a sort of dual functional proteins with specific binding to two distinct targets, which have become a focus of interest in antibody engineering and drug development research and have a promising future for wide applications in cancer immunotherapy and autoimmune disease. The key of clinical application and commercial-scale manufacturing of BsAbs is the amenability to assembly and purification of desired heterodimers. Advances in genetic engineering technology had resulted in the development of diverse BsAbs. Multiple recombinant strategies have been used to solve the mispairing problem between light and heavy chains, as well as to enforce accurate dimerization of heterologous heavy chains. There are 23 platforms available to generate 62 BsAbs which can be further divided into IgG-like ones and fragment-based ones, and more than 50 molecules are undergoing clinical trials currently. BsAbs with IgG-like architecture exhibit superior advantages in structure (similar to natural antibodies), pharmacokinetics, half-life, FcR-mediated function, and biological activity. This review considers various IgG-like BsAb generation approaches, summarizes the clinical applications of promising new BsAbs, and describes the mechanism of BsAbs in tumor therapy.

NuMat gets funds to build MOF plant

NuMat Technologies, one of C&EN’s 10 Start-Ups to Watch in 2016, has been awarded a $9 million contract from the US Army. NuMat will use the funds to build what it calls the first commercial-scale manufacturing plant for metal-organic frameworks (MOFs). A new lab will develop MOFs to protect soldiers from toxic agents. Their high surface area makes them better for adsorbing chemicals than materials currently used in gas masks and other protective gear. NuMat is also commercializing MOFs for electronics and gas storage.

Changing the ‘Art’ of Growing Cells into an Industrialized Platform for Commercial-Scale Manufacturing

Changing the ‘Art’ of Growing Cells into an Industrialized Platform for Commercial-Scale Manufacturing

Commercial Scale Manufacturing of Allogeneic Cell Therapy.

Allogeneic cell therapy products are generating encouraging clinical and pre-clinical results. Pluripotent stem cell (PSC) derived therapies, in particular, have substantial momentum and the potential to serve as treatments for a wide range of indications. Many of these therapies are also expected to have large market sizes and require cell doses of ≥109 cells. As therapeutic technologies mature, it is essential for the cell manufacturing industry to correspondingly develop to adequately support commercial scale production. To that end, there is much that can be learned and adapted from traditional manufacturing fields. In this review, we highlight key areas of allogeneic cell therapy manufacturing, identify current gaps, and discuss strategies for integrating new solutions. It is anticipated that cell therapy scale-up manufacturing solutions will need to generate batches of up to 2,000 L in single-use disposable formats, which constrains selection of currently available upstream hardware. Suitable downstream hardware is even more limited as processing solutions from the biopharmaceutical field are often not compatible with the unique requirements of cell therapy products. The advancement of therapeutic cell manufacturing processes to date has largely been developed with a cell biology driven approach, which is essential in early development. However, for truly robust and standardized production in a maturing field, a highly controlled manufacturing engineering strategy must be employed, with the implementation of automation, process monitoring and control to increase batch consistency and efficiency.

Laser Sintering of Liquid Metal Nanoparticles for Scalable Manufacturing of Soft and Flexible Electronics.

Soft, flexible, and stretchable electronics are needed to transmit power and information, and track dynamic poses in next-generation wearables, soft robots, and biocompatible devices. Liquid metal has emerged as a promising material for these applications due to its high conductivity and liquid phase state at room temperature; however, surface oxidation of liquid metal gives it unique behaviors that are often incompatible with scalable manufacturing techniques. This paper reports a rapid and scalable approach to fabricate soft and flexible electronics composed of liquid metal. Compared to other liquid metal patterning approaches, this approach has the advantages of compatibility with a variety of substrates, ease of scalability, and efficiency through automated processes. Nonconductive liquid metal nanoparticle films are sintered into electrically conductive patterns by use of a focused laser beam to rupture andablate particle oxide shells, and allow their liquid metal cores to escape and coalesce. The laser sintering phenomenon is investigated through comparison with focused ion beam sintering and by studying the effects of thermal propagation in sintered films. The effects of laser fluence, nanoparticle size, film thickness, and substrate material on resistance of the sintered films are evaluated. Several devices are fabricated to demonstrate the electrical stability of laser-patterned liquid metal traces under flexing, multilayer circuits, and intricately patterned circuits. This work merges the precision, consistency, and speed of laser manufacturing with the material benefits of liquid conductors on elastic substrates to demonstrate decisive progress toward commercial-scale manufacturing of soft electronics.

Industrializing Autologous Adoptive Immunotherapies: Manufacturing Advances and Challenges.

Cell therapy has proven to be a burgeoning field of investigation, evidenced by hundreds of clinical trials being conducted worldwide across a variety of cell types and indications. Many cell therapies have been shown to be efficacious in humans, such as modified T-cells and natural killer (NK) cells. Adoptive immunotherapy has shown the most promise in recent years, with particular emphasis on autologous cell sources. Chimeric Antigen Receptor (CAR)-based T-cell therapy targeting CD19-expressing B-cell leukemias has shown remarkable efficacy and reproducibility in numerous clinical trials. Recent marketing approval of Novartis' Kymriah™ (tisagenlecleucel) and Gilead/Kite's Yescarta™ (axicabtagene ciloleucel) by the FDA further underscores both the promise and legwork to be done if manufacturing processes are to become widely accessible. Further work is needed to standardize, automate, close, and scale production to bring down costs and democratize these and other cell therapies. Given the multiple processing steps involved, commercial-scale manufacturing of these therapies necessitates tighter control over process parameters. This focused review highlights some of the most recent advances used in the manufacturing of therapeutic immune cells, with a focus on T-cells. We summarize key unit operations and pain points around current manufacturing solutions. We also review emerging technologies, approaches and reagents used in cell isolation, activation, transduction, expansion, in-process analytics, harvest, cryopreservation and thaw, and conclude with a forward-look at future directions in the manufacture of adoptive immunotherapies.

Addressing the Challenges of Commercial-Scale Manufacture of Viral Vectors for CAR-T Therapies

Addressing the Challenges of Commercial-Scale Manufacture of Viral Vectors for CAR-T Therapies

Zein Nanoparticles and Strategies to Improve Colloidal Stability: A Mini-Review.

Zein, a protein extracted from maize, can be employed to easily produce nanoscale particles suitable for use as carrier systems. This review investigates the main methods for obtaining zein nanoparticles, as well as the problems and options available in the development of stable colloidal suspensions. Considerable gaps were identified in the literature concerning this topic, with studies being unclear about the factors that affect the stability of zein particles. In the vast majority of cases, no data are presented in relation to the stability of the formulations over time. It could be concluded that in order to produce a high quality system, detailed evaluation is required, considering factors including the zein concentration, pH, ionic strength, thermal treatment of the protein prior to preparation of the nanoparticles, strategies employing other materials as coatings, and the storage conditions. It is extremely important that these aspects should be considered during product development, prior to commercial-scale manufacture.

Biochemical analysis and the preliminary crystallographic characterization of d-tagatose 3-epimerase from Rhodobacter sphaeroides

Backgroundd-Tagatose 3-epimerase epimerizes d-fructose to yield d-psicose, which is a rare sugar that exists in small quantities in nature and is difficult to synthesize chemically. We aim to explore potential industrial biocatalysts for commercial-scale manufacture of this rare sugar. A d-tagatose 3-epimerase from Rhodobacter sphaeroides (RsDTE) has recently been identified as a d-tagatose 3-epimerase that can epimerize d-fructose to yield d-psicose with a high conversion rate.ResultsThe purified RsDTE by Ni-affinity chromatography, ionic exchange chromatography and gel filtration forms a tetramer in solution. The maximal activity was in Tris–HCl buffer pH 8.5, and the optimal temperature was at 35 °C. The product, d-psicose, was confirmed using HPLC and NMR. Crystals of RsDTE were obtained using crystal kits and further refined under crystallization conditions such as 10% PEG 8000,0.1 M HEPES pH 7.5, and 8% ethylene glycol at 20 °C using the sitting-drop vapor diffusion method. The RsDTE homology model showed that it possessed the characteristic TIM-barrel fold. Four residues, Glu156, Asp189, Gln215 and Glu250, forms a hydrogen bond network with the active Mn(II) for the hydride transfer reaction. These residues may constitute the catalytic tetrad of RsDTE. The residues around O1, O2 and O3 of the substrates were conserved. However, the binding-site residues are different at O4, O5 and O6. Arg118 formed the unique hydrogen bond with O4 of d-fructose which indicates RsDTE’s preference of d-fructose more than any other family enzymes.ConclusionsRsDTE possesses a different metal-binding site. Arg118, forming unique hydrogen bond with O4 of d-fructose, regulates the substrate recognition. The research on d-tagatose 3-epimerase or d-psicose 3-epimerase enzymes attracts enormous commercial interest and would be widely used for rare sugar production in the future.

Critically Evaluating the Benefits of Automation in Commercial-Scale Manufacture

Critically Evaluating the Benefits of Automation in Commercial-Scale Manufacture

Synthesis and Characterization of Potential Impurities of Canagliflozin

Multiple sources of anticipated process and degradation impurities of canagliflozin drug has been identified during its laboratory optimization and later in commercial scale manufacturing process. The structure of these impurities were confirmed by their independent synthesis as well as by their complete spectral characterization including 2D NMR, 19F NMR, and their chromatographic retention time profile. As a part of these investigations, three new impurities such as Hydroperoxide (Impurity-D), Sulfone (impurity-F) and Dimer (impurity-C) were characterized. The present work describe our detailed investigations on the structural characterization of canagliflozin impurities, and further their independent synthesis.

CordenPharma, GE form oligonucleotide services pact

CordenPharma International and GE Healthcare’s Dharmacon division are joining to provide what they call an end-to-end service for oligonucleotide-based drug discovery. The GE unit offers custom synthesis of complex oligos to support research. CordenPharma provides larger-scale oligos manufacturing, a service it launched last year after investing in a Boulder, Colo., plant it acquired from Roche in 2011. CordenPharma currently supports early-stage clinical development and expects to begin commercial-scale manufacturing by the end of the year.

Military may get mushroom buildings

Ecovative Design, a Green Island, N.Y.-based firm that develops structural materials out of organic matter bound with fungal mycelium, has been awarded up to $9.1 million from the U.S. Defense Advanced Research Projects Agency. The funds will support Ecovative’s work with researchers at Columbia University and MIT to develop self-repairing materials that can be used to grow structures in place, such as military bases in the field. The four-year project aims to demonstrate commercial-scale manufacturing.

Engineering Microneedle Patches for Vaccination and Drug Delivery to Skin

Microneedle patches (MNPs) contain arrays of solid needles measuring hundreds of microns in length that deliver drugs and vaccines into skin in a painless, easy-to-use manner. Optimal MNP design balances multiple interdependent parameters that determine mechanical strength, skin-insertion reliability, drug delivery efficiency, painlessness, manufacturability, and other features of MNPs that affect their performance. MNPs can be made by adapting various microfabrication technologies for delivery of small-molecule drugs, biologics, and vaccines targeted to the skin, which can have pharmacokinetic and immunologic advantages. A small number of human clinical trials, as well as a large and growing market for MNP products for cosmetics, indicate that MNPs can be used safely, efficaciously, and with strong patient acceptance. More advanced clinical trials and commercial-scale manufacturing will facilitate development of MNPs to realize their potential to dramatically increase patient access to otherwise-injectable drugs and to improve drug performance via skin delivery.

Glass Fibers: Quo Vadis?

Since the early 1930s, the process of melting glass and subsequently forming fibers, in particular discontinuous fiber glass or continuous glass filaments, evolved into commercial-scale manufacturing.[...]

Analysis of muscular nutritional composition in farmed Leuciscus spp. and their hybrids in a low saline-alkaline pond

PDF HTML阅读 XML下载 导出引用 引用提醒 低盐碱池塘养殖雅罗鱼及其杂交种肌肉营养成分分析 DOI: 作者: 作者单位: 1. 淡水鱼类育种国家地方联合工程实验室, 农业部淡水水产生物技术与遗传育种重点实验室, 中国水产科学研究院 特殊生境鱼类种质特性与抗逆育种重点实验室, 中国水产科学研究院 黑龙江水产研究所, 黑龙江 哈尔滨 150070;2. 上海海洋大学 水产与生命学院, 上海 201306 作者简介: 常玉梅(1978-),女,副研究员,专业方向为鱼类分子生物学.E-mail:changyumei@hrfri.ac.cn 通讯作者: 中图分类号: S96 基金项目: 国家自然科学基金项目（31461163004）；黑龙江省科技攻关项目（GCI2B307）. Analysis of muscular nutritional composition in farmed Leuciscus spp. and their hybrids in a low saline-alkaline pond Author: Affiliation: 1. National & Local United Engineering Laboratory of Freshwater Fish Breeding;Key Laboratory of Freshwater Aquatic Biotechnology and Genetic Breeding, Ministry of Agriculture;Key Laboratory of Fish Stress Resistance Breeding and Germplasm Characteristics in Special Habitats, Chinese Academy of Fishery Sciences;Heilongjiang River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Harbin 150070, China;2. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China Fund Project: 摘要 | 图/表 | 访问统计 | 参考文献 | 相似文献 | 引证文献 | 资源附件 | 文章评论 摘要:本研究旨在为池养雅罗鱼肌肉营养成分的评定和规模化配合饲料的配制提供基础数据。选取达里湖瓦氏雅罗鱼（L. idus，简称GT）以及种内杂交种（达里湖瓦氏雅罗鱼♀×松花江瓦氏雅罗鱼♂，简称DS）和种间杂交种（达里湖瓦氏雅罗鱼♀×高体雅罗鱼♂，简称DG）各15尾，每种鱼等量分成3组，测定其背部肌肉的一般营养成分、氨基酸和脂肪酸含量。结果显示：1）除水分含量无明显差异外，5种养殖雅罗鱼的灰分、粗蛋白和粗脂肪的含量均出现明显差异（<0.05）。其中DS杂交的灰分（1.77%）含量显著高于其他4种雅罗鱼（1.16%~1.22%），粗脂肪含量（7.23%）显著高于其双亲组合（DL 4.31%，SHJ 4.22%）及含量最低的DG杂交（3.32%），表现出超亲的杂种优势；5种养殖雅罗鱼具有较高的蛋白质含量（17.05%~18.80%），其中DG和DL的蛋白质含量明显高于其他3种养殖群体（<0.05），显示出较强的杂种优势；2）5种雅罗鱼肌肉中均测定出18种氨基酸，氨基酸总量（TAA）介于73.058%~79.289%，DG和DL的TAA含量、AAS、CS及EAAI指数均高于其他3种雅罗鱼，与其粗蛋白的测定结果相符，表明母本DL将其高蛋白的遗传特性很好地遗传给了子代，进一步证明了杂种优势；3）5种雅罗鱼肌肉中均检测到23种脂肪酸，饱和脂肪酸（SFA）、单不饱和脂肪酸（MUFA）和多不饱和脂肪酸（PUFA）含量分别为15.03%~19.06%、49.15%~53.92%和27.54%~29.40%，PUFA/SFA比值介于1.53~1.83；SFA和PUFA含量较高的是GT和DL，而MUFA含量较高的是DG和DS两个杂交种。综上所述，5种盐碱池塘养殖雅罗鱼的肌肉营养成分较均衡，属优质蛋白质；杂交选育可以有效改善杂交种的肌肉营养品质，获得的高脂高蛋白的DS杂交和低脂高蛋白的DG杂交可以满足不同的市场需求。 Abstract:This study was conducted to provide basic information for a comprehensive evaluation of nutritional components in the muscle tissues of five cultured fish species and to guide commercial-scale manufacturing of fish feed. Four-year-old originally from Dali Lake (DL) and Songhuajiang River progeny (SHJ), from the Ertix River (GT), and two hybrids, such as hybrids DS (DL♀×SHJ♂) and DG (DL♀×GT♂), were held in a low saline-alkaline pond. Fifteen fish from each genetic group were divided into three replicates, and their nutrient components and muscle amino acid and fatty acid contents were measured and analyzed. The results showed significant differences in ash, crude protein, and crude fat contents among the five genetic groups except that of water (<0.05). Ash content (1.77%) was significantly higher in DS than that in the other four genetic groups (1.16%-1.22%), and crude fat content (7.23%) was significantly higher than that of DL (4.31%), SHJ (4.22%), and the hybrid DG (3.32%). Eighteen amino acids were detected in the five genetic groups, and total amino acid contents were 73.06%-79.29%.The total amino acid components were significantly higher in DG and DL than those in the other three species (<0.05). In addition, the DG and DL groups had higher amino acid, chemical, and essential amino acid index scores than those of the others, which coincided with the higher crude protein content in DL, indicating that the higher crude protein in DG was inherited from DL, and was evidence of hybrid vigor. Twenty-three fatty acids were detected in the five genetic groups, and the saturated fatty acid (SFA), mono-unsaturated fatty acid (MUFA), and polyunsaturated fatty acid (PUFA) contents were 15.03%-19.06%, 49.15%-53.92%, and 27.54%-29.40%, respectively. The PUFA to SFA ratio was 1.53-1.83. GT and DL had relatively higher SFA and PUFA contents, and the DG and DS hybrids had higher MUFA contents. Taken together, the five cultured genetic groups had balanced nutrient components in muscle and could be excellent protein sources for human consumption. Nutrient quality was improved effectively through hybridization; thus, satisfying market requirements with higher lipid and protein contents from DS and lower lipid but higher protein contents from DG. 参考文献 相似文献 引证文献

Design and Comparison of Tubular and Pipes-in-Series Continuous Reactors for Direct Asymmetric Reductive Amination

A high pressure, high temperature direct asymmetric reductive amination (DARA) reaction was run in three different continuous reactor types; coiled tubes, horizontal pipes in series, and vertical pipes in series. The goal of this work was to design a continuous reactor scalable to large volumes needed for commercial scale manufacturing. With about 4000:1 substrate to dissolved catalyst ratio, the DARA reaction achieved about 90% conversion to desired product in 12 h, about 8% conversion to undesired ketone, and about 95% ee, in batch and in all 3 types of continuous reactors. The 3 continuous reactors were compared on the basis of vapor/liquid mass transfer rates, overall pressure drop, and axial dispersion. Vertical pipes in series were selected as the best option because of scalability and vapor/liquid mass transfer rates, although horizontal pipes had lower pressure drop and less potential for surging.