Scalable Manufacturing Process Research Articles

Nanoscale Strainability of Graphene by Laser Shock-Induced Three-Dimensional Shaping

Graphene has many promising physical properties. It has been discovered that local strain in a graphene sheet can alter its conducting properties and transport gaps. It is of great importance to develop scalable strain engineering techniques to control the local strains in graphene and understand the limit of the strains. Here, we present a scalable manufacturing process to generate three-dimensional (3D) nanostructures and thus induce local strains in the graphene sheet. This process utilizes laser-induced shock pressure to generate 3D tunable straining in the graphene sheet. The size dependent straining limit of the graphene and the critical breaking pressure are both studied. It is found that the graphene film can be formed to a circular mold (∼50 nm in diameter) with an aspect ratio of 0.25 and strain of 12%, and the critical breaking pressure is 1.77 GPa. These values were found to be decreasing with the increase of mold size. The local straining and breaking of graphene film are verified by Raman spectra. Large scale processing of the graphene sheet into nanoscale patterns is presented. The process could be scaled up to roll-to-roll process by changing laser beam size and scanning speed. The presented laser shock straining approach is a fast, tunable, and low-cost technique to realize strain engineering of graphene for its applications in nanoelectrical devices.

Micro and nano patterning of carbon electrodes for bioMEMS

Carbon based bioMEMS are an emerging class of miniaturized biomedical devices. Due to the numerous advantages such as scalable manufacturing processes, inexpensive and readily available precursor polymer materials, tunable surface properties and biocompatibility, carbon has become a preferred material for a wide variety of future biosensing applications. In this article, we review fabrication methodologies for carbon-MEMS (CMEMS) and carbon-NEMS (CNEMS) devices and strategies for their surface modification for biocompatibility and for biosensing. We also discuss the underlying graphitic and glassy microstructures of carbon since the biocompatibility of carbon and its microstructures are intertwined. We conclude by exploring various CMEMS and CNEMS devices that have been successfully fabricated.

Ideal Energy‐Level Alignment at the ZnO/P3HT Photovoltaic Interface

AbstractHybrid semiconductor‐polymer nanostructured solar cells hold the promise of photovoltaic energy conversion based on abundant and nontoxic materials and scalable manufacturing processes. After a decade of intense research activity, hybrid solar cells still exhibit low short‐circuit currents and moderate open‐circuit voltages. These bottlenecks call for a detailed understanding of the physics underlying the device operation at the nanoscale. Using first‐principles calculations the ideal energy‐level alignment of hybrid solar cell interfaces based on the wide bandgap semiconductor ZnO and the polymer poly(3‐hexylthiophene) (P3HT) is investigated. The interfacial charge transfer is quantified and it is shown that this effect increases the ideal open‐circuit voltage with respect to the electron‐affinity rule by as much as 0.5 V. The results of this work suggests that there is significant room for optimizing this class of excitonic solar cells by tailoring the semiconductor/polymer interface at the nanoscale.

Gastroresistant capsular device prepared by injection molding

In the present work, the possibility of manufacturing by injection molding (IM) a gastro-resistant capsular device based on hydroxypropyl methyl cellulose acetate succinate (HPMCAS) was investigated. By performing as an enteric soluble container, such a device may provide a basis for the development of advantageous alternatives to coated dosage forms. Preliminarily, the processability of the selected thermoplastic polymer was evaluated, and the need for a plasticizer (polyethylene glycol 1500) in order to counterbalance the glassy nature of the molded items was assessed. However, some critical issues related to the physical/mechanical stability (shrinkage and warpage) and opening time of the device after the pH change were highlighted. Accordingly, an in-depth formulation study was carried out taking into account differing release modifiers potentially useful for enhancing the dissolution/disintegration rate of the capsular device at intestinal pH values. Capsule prototypes with thickness of 600 and 900 μm containing Kollicoat(®) IR and/or Explotab(®) CLV could be manufactured, and a promising performance was achieved with appropriate gastric resistance in pH 1.2 medium and break-up in pH 6.8 within 1h. These results would support the design of a dedicated mold for the development of a scalable manufacturing process.

A Scalable System for Production of Functional Pancreatic Progenitors from Human Embryonic Stem Cells

Development of a human embryonic stem cell (hESC)-based therapy for type 1 diabetes will require the translation of proof-of-principle concepts into a scalable, controlled, and regulated cell manufacturing process. We have previously demonstrated that hESC can be directed to differentiate into pancreatic progenitors that mature into functional glucose-responsive, insulin-secreting cells in vivo. In this study we describe hESC expansion and banking methods and a suspension-based differentiation system, which together underpin an integrated scalable manufacturing process for producing pancreatic progenitors. This system has been optimized for the CyT49 cell line. Accordingly, qualified large-scale single-cell master and working cGMP cell banks of CyT49 have been generated to provide a virtually unlimited starting resource for manufacturing. Upon thaw from these banks, we expanded CyT49 for two weeks in an adherent culture format that achieves 50–100 fold expansion per week. Undifferentiated CyT49 were then aggregated into clusters in dynamic rotational suspension culture, followed by differentiation en masse for two weeks with a four-stage protocol. Numerous scaled differentiation runs generated reproducible and defined population compositions highly enriched for pancreatic cell lineages, as shown by examining mRNA expression at each stage of differentiation and flow cytometry of the final population. Islet-like tissue containing glucose-responsive, insulin-secreting cells was generated upon implantation into mice. By four- to five-months post-engraftment, mature neo-pancreatic tissue was sufficient to protect against streptozotocin (STZ)-induced hyperglycemia. In summary, we have developed a tractable manufacturing process for the generation of functional pancreatic progenitors from hESC on a scale amenable to clinical entry.

Development of a Large-Scale Synthetic Route to Manufacture (−)-Huperzine A

A safe, practical and scalable process for manufacture of (−)-huperzine A has been developed and scaled up to manufacture several hundred grams of (−)-huperzine A with chemical and optical purity of >99%. The process consists of 11 chemical stages starting from commercially available materials with only nine isolation steps and no chromatography purification. This process provides a reliable and cost-effective source of synthetic (−)-huperzine A and its derivatives for pharmaceutical and nutraceutical markets.

Developing Assays to Address Identity, Potency, Purity and Safety: Cell Characterization in Cell Therapy Process Development

A major challenge to commercializing cell-based therapies is developing scalable manufacturing processes while maintaining the critical quality parameters (identity, potency, purity, safety) of the final live cell product. Process development activities such as extended passaging and serum reduction/elimination can facilitate the streamlining of cell manufacturing process as long as the biological functions of the product remain intact. Best practices in process development will be dependent on cell characterization; a thorough understanding of the cell-based product. Unique biological properties associated with different types of cell-based products are discussed. Cell characterization may be used as a tool for successful process development activities, which can promote a candidate cell therapy product through clinical development and ultimately to a commercialized product.

The significance of plasmid DNA preparations contaminated with bacterial genomic DNA on inflammatory responses following delivery of lipoplexes to the murine lung

Non-viral gene transfer using plasmid DNA (pDNA) is generally acknowledged as safe and non-immunogenic compared with the use of viral vectors. However, pre-clinical and clinical studies have shown that non-viral (lipoplex) gene transfer to the lung can provoke a mild, acute inflammatory response, which is thought to be, partly, due to unmethylated CG dinucleotides (CpGs) present in the pDNA sequence. Using a murine model of lung gene transfer, bronchoalveolar lavage fluid was collected following plasmid delivery and a range of inflammatory markers was analysed. The results showed that a Th1-related inflammatory cytokine response was present that was substantially reduced, though not abolished, by using CpG-free pDNA. The remaining minor level of inflammation was dependent on the quality of the pDNA preparation, specifically the quantity of contaminating bacterial genomic DNA, also a source of CpGs. Successful modification of a scalable plasmid manufacturing process, suitable for the production of clinical grade pDNA, produced highly pure plasmid preparations with reduced genomic DNA contamination. These studies help define the acceptable limit of genomic DNA contamination that will impact FDA/EMEA regulatory guidelines defining clinical grade purity of plasmid DNA for human use in gene therapy and vaccination studies.

Binder free porous ultrafine/nano structured LiCoO2 cathode from plasma deposited cobalt

Increasing global demand for rechargeable lithium (Li) ion batteries has been driving the research on innovative processing techniques and material systems for cheaper production of battery electrodes. While cost reduction is important, obtaining desired microstructures in the active electrode material is also very critical for efficient performance of the batteries. Conventional processing of bulk scale Li-ion battery electrodes involves time consuming and multi-step processes starting from the production of active powder materials, blending with conductive additives and binders to develop the electrode material coating on a current collector. On the other hand, thin film battery technologies employ expensive vapor based or sputtering or laser ablation techniques to develop the electrodes. In this study, an innovative, rapid and a two step scalable manufacturing process has been developed. While capable of developing porous and ultrafine/nano structured oxide based LiCoO2 cathode material directly on a charge collector from metallic Cobalt (Co) coatings, the process does not require polymeric binders. Following this approach, LiCoO2 cathodes were synthesized directly on a stainless steel charge collector from plasma sprayed Co coatings via thermal treatments using aqueous LiNO3 solution. X-ray diffraction (XRD) studies confirmed presence of LiCoO2 hexagonal phase. Microstructural and phase analysis showed porous active material with ultrafine/nano structural features along with imperfections (e.g., dislocations). Electrochemical characterization illustrated an average voltage around 3.9V with a specific discharge capacity around 70–85% of the nominal capacity (∼138mAh/g) against Li counter electrode. However, process optimization in terms of plasma spray coatings and thermal treatments, and addition of carbon may enhance the performance of LiCoO2 electrodes. Absence of polymeric binders makes these electrodes suitable for high temperature battery applications.

Scale-up and manufacturing of clinical-grade self-inactivating γ-retroviral vectors by transient transfection.

The need for γ-retroviral (gRV) vectors with a self-inactivating (SIN) design for clinical application has prompted a shift in methodology of vector manufacturing from the traditional use of stable producer lines to transient transfection-based techniques. Herein, we set out to define and optimize a scalable manufacturing process for the production of gRV vectors using transfection in a closed-system bioreactor in compliance with current good manufacturing practices (cGMP). The process was based on transient transfection of 293T cells on Fibra-Cel disks in the Wave Bioreactor. Cells were harvested from tissue culture flasks and transferred to the bioreactor containing Fibra-Cel in the presence of vector plasmid, packaging plasmids and calcium-phosphate in Dulbecco's modified Eagle's medium and 10% fetal bovine serum. Virus supernatant was harvested at 10-14 h intervals. Using optimized procedures, a total of five ecotropic cGMP-grade gRV vectors were produced (9 liters each) with titers up to 3.6 × 10(7) infectious units per milliliter on 3T3 cells. One GMP preparation of vector-like particles was also produced. These results describe an optimized process for the generation of SIN viral vectors by transfection using a disposable platform that allows for the generation of clinical-grade viral vectors without the need for cleaning validation in a cost-effective manner.

Enabling a robust scalable manufacturing process for therapeutic exosomes through oncogenic immortalization of human ESC-derived MSCs

BackgroundExosomes or secreted bi-lipid vesicles from human ESC-derived mesenchymal stem cells (hESC-MSCs) have been shown to reduce myocardial ischemia/reperfusion injury in animal models. However, as hESC-MSCs are not infinitely expansible, large scale production of these exosomes would require replenishment of hESC-MSC through derivation from hESCs and incur recurring costs for testing and validation of each new batch. Our aim was therefore to investigate if MYC immortalization of hESC-MSC would circumvent this constraint without compromising the production of therapeutically efficacious exosomes.MethodsThe hESC-MSCs were transfected by lentivirus carrying a MYC gene. The transformed cells were analyzed for MYC transgene integration, transcript and protein levels, and surface markers, rate of cell cycling, telomerase activity, karyotype, genome-wide gene expression and differentiation potential. The exosomes were isolated by HPLC fractionation and tested in a mouse model of myocardial ischemia/reperfusion injury, and infarct sizes were further assessed by using Evans' blue dye injection and TTC staining.ResultsMYC-transformed MSCs largely resembled the parental hESC-MSCs with major differences being reduced plastic adherence, faster growth, failure to senesce, increased MYC protein expression, and loss of in vitro adipogenic potential that technically rendered the transformed cells as non-MSCs. Unexpectedly, exosomes from MYC-transformed MSCs were able to reduce relative infarct size in a mouse model of myocardial ischemia/reperfusion injury indicating that the capacity for producing therapeutic exosomes was preserved.ConclusionOur results demonstrated that MYC transformation is a practical strategy in ensuring an infinite supply of cells for the production of exosomes in the milligram range as either therapeutic agents or delivery vehicles. In addition, the increased proliferative rate by MYC transformation reduces the time for cell production and thereby reduces production costs.

Sustained Enzymatic Correction by rAAV-Mediated Liver Gene Therapy Protects Against Induced Motor Neuropathy in Acute Porphyria Mice

Acute intermittent porphyria (AIP) is characterized by a hereditary deficiency of hepatic porphobilinogen deaminase (PBGD) activity. Clinical features are acute neurovisceral attacks accompanied by overproduction of porphyrin precursors in the liver. Recurrent life-threatening attacks can be cured only by liver transplantation. We developed recombinant adeno-associated virus (rAAV) vectors expressing human PBGD protein driven by a liver-specific promoter to provide sustained protection against induced attacks in a predictive model for AIP. Phenobarbital injections in AIP mice induced porphyrin precursor accumulation, functional block of nerve conduction, and progressive loss of large-caliber axons in the sciatic nerve. Hepatocyte transduction showed no gender variation after rAAV2/8 injection, while rAAV2/5 showed lower transduction efficiency in females than males. Full protection against induced phenobarbital-attacks was achieved in animals showing over 10% of hepatocytes expressing high amounts of PBGD. More importantly, sustained hepatic expression of hPBGD protected against loss of large-caliber axons in the sciatic nerve and disturbances in nerve conduction velocity as induced by recurrent phenobarbital administrations. These data show for the first time that porphyrin precursors generated in the liver interfere with motor function. rAAV2/5-hPBGD vector can be produced in sufficient quantity for an intended gene therapy trial in patients with recurrent life-threatening porphyria attacks.

A flexible OLED display with an OTFT backplane made by scalable manufacturing process

Abstract— A full‐color top‐emission AMOLED display driven by an organic TFT backplane manufactured using a scalable, lift‐off‐free, and shadow‐mask‐free process has been developed. It was shown that cost‐efficient copper can be used for S/D electrodes. The display has shown no significant degradation over a storage life of more than 10 months and operation over 25 hours during which the display is bent over 10,000 times.

A rapid method for creating drug implants: Translating laboratory‐based methods into a scalable manufacturing process

Low compliance with medication is the major cause of poor outcome in schizophrenia treatment. While surgically implantable solvent-cast pellets were produced to improve outcome by increased compliance with medication, this process is laborious and time-consuming, inhibiting its broader application (Siegel et al., Eur J Pharm Biopharm 2006;64:287-293). In this study, the previous fabrication process was translated to a continuous and scalable extrusion method. Extrusion processes were modified based on in vitro release studies, drug load consistency examination, and surface morphology analysis using scanning electron microscopy. Afterward, optimized haloperidol implants were implanted into rats for preliminary analysis of biocompatibility. Barrel temperature, screw speed and resulting processing pressure influenced surface morphology and drug release. Data suggest that fewer surface pores shift the mechanism from bulk to surface PLGA degradation and longer lag period. Results demonstrate that extrusion is a viable process for manufacturing antipsychotic implants.

Visible frequency magnetic activity in silver nanocluster metamaterial

We experimentally observe magnetic resonance in the visible frequency region from self-assembled silver nanocluster metamaterials. Extensive numerical modeling studies were conducted to find the optimal nanocluster dimensions. Self-assembly of silver nanoparticles coated with nanoscale silica coating was then performed on polymer templates fabricated by laser interference lithography. The nanoclusters supported magnetic resonance in the visible region, and the extracted effective permeability exhibited Lorentz-like resonance. The experimentally observed lowest value for the real part of permeability was 0.06. The nanocluster metamaterial represents a practical metamaterial architecture that is compatible with the scalable bottom-up manufacturing process.

High Critical Current YBCO Films Prepared by an MOD Process on RABiTS Templates

The metal organic deposition (MOD) of YBCO high temperature superconducting films on RABiTS (rolling assisted biaxially textured substrates) templates has been developed at American Superconductor as a low-cost, scalable manufacturing process for the commercialization of the second generation (2G) HTS wire. The MOD process is based on the deposition of a triflu-oroacetate (TFA) based metal organic precursor film which is converted, in an ex-situ process, to the superconducting YBCO film. A major goal of the development has been achieving high critical currents. This paper reports the preparation and characterization of MOD-YBCO films with critical currents exceeding 500 A/cm-w (77 K, self-field) using a scaleable thick film approach on RABiTS templates. The high critical current films were obtained through optimization of the precursor composition, nucleation and growth conditions. The through-thickness dependence of the critical current density of MOD Alms as a function of film thickness and a correlation of the through-thickness transport properties and microstructure of the thick MOD/RABiTS samples is reported.

7.4: A Flexible Plastic SVGA e-Paper Display

We have developed a scalable manufacturing process for active matrix displays compatible with low temperature plastic substrates and have applied this technology to the fabrication of flexible SVGA display backplanes. We combined these backplanes with E Ink® Imaging Film to produce 100 PPI SVGA (800×600 pixel) displays exhibiting grey scale and with a high aperture ratio. Power consumption is zero in between image changes. Flexible high information content e-paper displays will change the way in which information is conveyed by enabling lightweight, robust e-reader devices.

P-65: Developing a Scalable and Adaptable Ink Jet Printing Process for OLED Displays

The feasibility of using ink jet printing to define hole conduction and electroluminescent layers in OLED display has been well demonstrated. The particular motivation for ink jet printing has been driven by the prospect of developing scalable and adaptable manufacturing processes, enabling large substrate sizes to be processed, without the requirement for expensive product specific tooling. In this paper we discuss the implication of scalable and adaptable criteria for an ink jet print process and demonstrate how this can be achieved by development of suitable ink formulations.

A scalable manufacturing process for flexible active-matrix e-paper displays

Journal of the Society for Information DisplayVolume 8, Issue 1 p. 1-1 Introduction Andras I. Lakatos, Andras I. Lakatos EditorSearch for more papers by this author Andras I. Lakatos, Andras I. Lakatos EditorSearch for more papers by this author First published: 18 June 2012 https://doi.org/10.1889/1.1828693Citations: 11AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article.Citing Literature Volume8, Issue1March 2000Pages 1-1 RelatedInformation

Immobilized cobalt affinity chromatography provides a novel, efficient method for herpes simplex virus type 1 gene vector purification.

Herpes simplex virus type 1 (HSV-1) is a promising vector for gene therapy applications, particularly at peripheral nerves, the natural site of virus latency. Many gene vectors require large particle numbers for even early-phase clinical trials, emphasizing the need for high-yield, scalable manufacturing processes that result in virus preparations that are nearly free of cellular DNA and protein contaminants. HSV-1 is an enveloped virus that requires the development of gentle purification methods. Ideally, such methods should avoid centrifugation and may employ selective purification processes that rely on the recognition of a unique envelope surface chemistry. Here we describe a novel method that fulfills these criteria. An immobilized metal affinity chromatography (IMAC) method was developed for the selective purification of vectors engineered to display a high-affinity binding peptide. Feasibility studies involving various transition metal ions (Cu2+, Zn2+, Ni2+, and Co2+) showed that cobalt had the most desirable features, which include a low level of interaction with either the normal virus envelope or contaminating DNA and proteins. The introduction of a cobalt-specific recognition element into the virus envelope may provide a suitable target for cobalt-dependent purification. To test this possibility, we engineered a peptide with affinity for immobilized cobalt in frame in the heparan sulfate binding domain of HSV-1 glycoprotein B, which is known to be exposed on the surface of the virion particle and recombined into the viral genome. By optimizing the IMAC loading conditions and reducing cobalt ion leakage, we recovered 78% of the tagged HSV-1 recombinant virus, with a >96% reduction in contaminating proteins and DNA.