Rapid Manufacturing Process Research Articles

A Novel Machine-Learning-Based Procedure to Determine the Surface Finish Quality of Titanium Alloy Parts Obtained by Heat Assisted Single Point Incremental Forming

Single point incremental forming (SPIF) is a cheap and flexible sheet metal forming process for rapid manufacturing of complex geometries. Additionally, it is important for engineers to measure the surface finish of work pieces to assess their quality and performance. In this paper, a predictive model based on machine learning and computer vision was developed to estimate arithmetic mean surface roughness (Ra) and maximum peak to valley height (Rz) of Ti6Al4V parts obtained by SPIF. An image database was prepared to train different classification algorithms in accordance with a supervised learning approach. A speeded up robust feature (SURF) detector was used to obtain visual vocabulary so that the classifiers are able to group the photographs into classes. The experimental results indicated that the proposed predictive method shows great potential to determine the surface quality, as classifiers based on a support vector machine with a polynomial kernel are suitable for this purpose.

Rapid manufacturing of micro-drilling devices using FFF-type 3D printing technology

Micro-drilling devices with different blade shapes were fabricated with a rapid and facile manufacturing process using three-dimensional (3D) printing technology. The 3D-printed casting mold was utilized to customize the continuous shape of the blades without the need for expensive manufacturing tools. A computational fluid dynamics simulation was performed to estimate the pressure differences (fluidic resistance) around each rotating device in a flowing stream. Three types of blades (i.e., 45°, 0°, and helical type) were manufactured and compared to a device without blades (i.e., plain type). As a result, the device with the 45° blades exhibited the best drilling performance. At a rotational speed of 1000 rpm, the average drilling depth of the device with the 45° blades to penetrate artificial thrombus for 90 s was 3.64 mm, which was ~ 2.4 times longer than that of helical blades (1.51 mm). This study demonstrates the feasibility of using 3D printing to fabricate microscale drilling devices with sharp blades for various applications, such as in vivo microsurgery and clogged water supply tube maintenance.

Influence of Automated Fiber Placement Parameters on Thermoplastic Composite Blanks Used on Stamp Forming Process

In recent years, advanced manufacturing processes have been developed to increase the speed of production in order to reduce production costs. At the scale of thermoplastic composites, the translation is the combination of advanced manufacturing processes. The focus in this study is more specifically on the coupling of automated lay-up (AFP) and stamp forming processes. To date, a consolidation process, such as press-consolidation of thermoplastic composites, obtained blanks. Several trials have begun using an automated fiber placement consolidation to reduce manufacturing time and use unidirectional material. However, the combination of AFP and stamp forming is useful if it optimizes this process without the blank’s full consolidation, which by resulting reduces the manufacturing time. This study estimates blank characteristics through thermal history imposed by a more rapid manufacturing process. A set of blanks with varying process parameters is produced to investigate the influence at the microscopic scale. The interface behaviour is observed with optical microscope and image processing. A statistical study applied to the process is carried out in order to relate the material observations to the input parameters. The results of this study are used for the study of the next process of the combination: the stamp forming.

A Novel Rapid Manufacturing Process for Metal Lattice Structure.

A novel lattice structure manufacturing process is proposed in this article, which has the potential to overcome the manufacturing shortcomings of small-scale metal lattice structure. The proposed hierarchical process has four segments: Design, Bending, Dip, and Join (DBDJ). The proposed research use one-dimensional metallic wires/rods instead of powder, two-dimensional sheet, or liquid metal, which is highly transformative than the status quo. The topology-based design technique is focused to construct the lattice structure using a continuous thin rod. The layers are stacked in an additive manner to construct the three-dimensional lattice structure. The dip-coating meditate material transfer facilitates the node joining using transient liquid phase diffusion bonding, and hence, the manufacturing of the complex lattice structure is performed. The research framework provides a unique and holistic approach from design to manufacturing for realizing small-scale metallic lattice structure. A range of multiscale lattice structure is manufactured with the proposed DBDJ process. Very low relative density (∼3.8%) unit cell is achieved, and compressive tests demonstrate no failure at the joining node, which is reported in this article.

Investigation of mechanical properties and form error of the components fabricated by rapid prototyping: A review

3D printing is emerging as one of the most important and advanced technique of manufacturing. Nowadays, manufacturing industries is focusing on reduction of lead time of product development and cost of product. 3D printing has large application in many fields such as aerospace and defence, medical, consumer goods etc .The purpose of this study was to present a concise review on rapid manufacturing processes. This paper includes historical development, available technologies, materials and their tangible benefits. Researchers have analyzes the influence of process parameters like orientation, layer thickness, air gap, printing direction etc. on properties of components which are fabricated through 3D printing. The noble contribution of this paper is to present systematic approach to enable potential for academics user as well as industry persons to select appropriate technology.

Feasibility analysis of manufacturing using rapid prototyping: A review

Nowadays manufacturing industries is focusing on reduction of lead time of product development and cost of product. Rapid prototyping has become a solution to manufacturing industry globally. The purpose of this study is to present a concise review on rapid manufacturing processes. This paper includes historical development, available technologies, materials and their tangible benefits. The noble contribution of this paper is to present systematic approach to enable potential for academics user as well as industry persons to select appropriate technology.

Effect of specimen parameters on mixed-mode I/II stress intensity factors for additive manufactured slant edge crack plate

Abstract Product development using rapid prototyping methods for direct application as end product, by virtue of advances in materials and technology, is new emerging concept in manufacturing. These products exhibit inherent cracks due to either the process of building or post processing which make it necessary to study its fracture aspects. In current study mixed-mode I/II stress intensity factors are investigated for an inclined crack in edge crack specimen, built with rapid manufacturing process employing Polyjet technology. The parameters in consideration are crack length ratio, crack angle and layer-built angle. It is observed that the specimen parameters under consideration affect significantly the mode I and mode II stress intensity factors values of slant edge crack plate.

Effects of solution-aging treatments on microstructure features, mechanical properties and damage behaviors of additive manufactured Ti–6Al–4V alloy

Hot isostatic pressing (HIPing) treatment was generally carried out to eliminate possible defects and to homogenize the microstructure for Ti–6Al–4V alloy fabricated by the Electron beam rapid manufacturing (EBRM) process. But the HIPed alloy with relative coarse lamellar microstructure exhibited a low strength compared to the as-built one. Post solution-aging treatments were necessary to optimize the HIPed alloy's tensile properties. So far, the relationship between the heat-treated microstructure features and corresponding mechanical properties of EBRM-produced Ti–6Al–4V has not been completely understood. Therefore, in the present work, solution-aging treatments under various solution temperatures (930 °C–990 °C) were applied to the HIPed Ti–6Al–4V produced by the EBRM, and the corresponding microstructures and mechanical properties were investigated. The results showed that solution-aging treatments could increase the HIPed alloy's tensile strength but decrease its ductility. With increasing solution temperature, the tensile strength presented an upward tendency until to 980 °C, whereas a decrease after 990 °C solution treatment. The variation in tensile properties was caused by the microstructures evolved with solution temperatures. In-situ tensile results showed that the deformation and damage behaviors of the alloy were also highly affected by the solution temperatures. Owning to a decrease of content of the αp with increase of solution temperature, the alloy at high solution temperature presented superior resistance of crack initiation but inferior resistance of crack growth than that at low solution temperature.

Influence of hatch spacing and laser spot overlapping on heat transfer during laser powder bed fusion of aluminum alloy

Direct metal laser sintering is a kind of laser powder bed fusion process that emerged as an advanced manufacturing process in the recent era of industry 4.0. This is a rapid manufacturing process where repeated melting and solidification takes place during scanning of the powder bed. The characteristics of the as-built component depend on the melting and solidification behavior which is directly influenced by the processing conditions. Intending to make a superior part in this process, it is indispensable to comprehend the impact of process parameters on temperature distribution, melting and solidification behavior, etc. In the present study, the authors developed an improved three-dimensional transient heat transfer model and studied the influence of hatching space and laser spot overlapping on temperature distribution with molten pool behavior on the AlSi10Mg powder bed. From this investigation, it was observed that the temperature on the AlSi10Mg powder bed decreases with an increase in hatch spaces and conversely with increasing the laser spot overlapping the temperature increases. Also, the molten pool dimension changes with variation of hatch spacing and laser spot overlapping. By combining these studies, the manufactures can able to optimize the hatch space and laser spot overlap along with other processing parameters to fabricate quality components.

Indium tin oxide nanowires manufactured via printing and laser irradiation

Metallic and semiconductor nanowires can provide dramatically increased electrical and optical properties in a wide range of fields, ranging from photovoltaics to sensors and catalysts. In this research, a rapid manufacturing process has been developed for printing indium tin oxide microparticles and converting them into nanowires. Microparticle indium tin oxide (ITO) inks were formulated and printed. These were then converted into hierarchical nanowire films via laser irradiation (980 nm, NIR) with raster speeds of 40 mm s−1 in air, much faster compared to traditional manufacturing processes. For a 4 cm2 film, only 40 s of processing were required. A full materials characterization was performed on the materials pre and post laser processing with the most probable conversion mechanism found to be a laser induced carbothermal reduction process. Microstructural, chemical, and crystallographic evidence of the laser induced carbothermal reduction process were derived from SEM, XRD, XPS and TEM analysis. Compared to conventionally heat-treated printed samples, laser processing was found to increase the conductivity of the printed ITO from 0.88% to 40.47% bulk conductivity. This research demonstrates the ability of printing and laser processing to form nanowires in a high-speed manufacturing context, thereby enabling the development of printed non-transparent ITO nanowire electronics and devices.

Rapid response platform to manufacture human immunoglobulins

Abstract A rapidly deployable drug product for use in the early stages of a disease outbreak could prevent further spread of the pathogen. Emergent BioSolutions is developing a rapid response platform (RRP) to manufacture human immunoglobulins (HIG) for use as a passive immunotherapy during public health emergencies. Passive immunotherapies derived from either convalescent plasma or purified immunoglobulins (Ig) have a long history of success in treating a range of viral, bacterial and toxin diseases, as they provide the immediate benefit of a protective response. Emergent's RRP is being developed to address the need for front-line therapy/prophylaxis with immediate benefit with the potential to overcome the current limitations of convalescent plasma. Emergent's proposed RRP begins with identification of plasma donors with measurable levels of pathogen-specific antibodies using a field deployable real-time donor screening assay. Plasma collected from eligible donors is treated using the Mirasol® Pathogen Reduction Technology, pooled, purified and concentrated into HIG using Emergent's rapid HIG manufacturing process to produce product formulated for intramuscular administration. Our rapid HIG process is chromatography based and each 30 L run aims to purify, formulate, and fill ∼65 doses (1 g of total IgG) with testing and release onsite for immediate use. Multiple innovations to streamline operations, including single-use technologies and minimal human interfaces, allow for the manufacture of HIG product in ∼30 hours. To house the process, Emergent has designed and constructed a working modular manufacturing unit (MMU) prototype. The MMU consists of a modified 53-foot shipping container designed for multiple modes of transport, to meet ISO 8 environment requirements with self-contained utilities, including HVAC, and can run off a diesel generator or local power. Product from multiple development runs within the MMU have been characterized and validation work is ongoing. Key messages A modular manufacturing unit is being developed with the potential to provide a local capability to produce health solutions for endemic diseases. A modular manufacturing unit is being developed with the potential to rapidly manufacture human immunoglobulins during public health emergencies and pandemics.

Electric discharge machining using rapid manufactured complex shape copper electrode with cryogenic cooling channel

In this study, a rapid manufacturing process based on the combination of polymer three-dimensional printing and pressureless loose sintering was explored for the fabrication of complex shape electric discharge machining pure copper electrodes with the cryogenic cooling channel. The fabricated electrodes were used to perform electric discharge machining on D2 steel workpiece. The comparative study was performed on material removal and electrode wear rates between the solid copper electrode, rapid manufactured electrode without cryogenic cooling and with cryogenic cooling. Also, the surface characteristics of the worn electrode and the machined workpiece were studied with and without cryogenic cooling. The significant effect of the cryogenic cooling on the electrode wear rate and the surface roughness was observed. Better surface finish, small cracks and less debris were notified on the workpiece surface machined with rapid manufactured electrode with cryogenic cooling due to rapid dissipation of the heat from the surface of the electrode after machining. Similarly, few cracks and low carbon deposition was observed on the rapid manufactured electrode with cryogenic cooling surface after machining as compared to rapid manufactured electrode without cryogenic cooling. The sharp corner edges of the complex shape tool in rapid manufactured electrode with cryogenic cooling were retained after machining due to low melting and vaporization of the electrode material. The dimensional deviation of the machined surface with respect to computer-aided design model was compared. The rapid manufactured electrode with cryogenic cooling was found to machine the more accurate complex shape features in terms of dimensions on the workpiece as compared to rapid manufactured electrode without cryogenic cooling.

Abstract 6593: PRGN-3005 UltraCAR-T™: multigenic CAR-T cells generated using non-viral gene delivery and rapid manufacturing process for the treatment of ovarian cancer

Abstract Ovarian cancer is the most lethal of gynecologic cancers. Chimeric antigen receptor (CAR)-T cells have improved clinical outcomes in hematological malignancies; however similar responses have not yet been achieved for solid tumors. Furthermore, traditional methods for CAR-T manufacturing utilize viral vectors and weeks of ex vivo expansion to achieve clinically relevant cell numbers. This leads to CAR-T cells with an exhausted T cell phenotype, high manufacturing costs, and treatment delays. We have developed the UltraCAR-T™ platform to overcome these limitations by utilizing an advanced non-viral gene delivery system and a rapid, decentralized manufacturing process. Our non-viral gene delivery platform is based on Sleeping Beauty transposon system precision engineered for optimal delivery of multiple genes of UltraCAR-T. UltraCAR-T cells are manufactured at medical center's cGMP facility using autologous T cells and administered back to patient only one day after gene transfer. PRGN-3005 UltraCAR-T cells simultaneously co-express (i) a CAR specifically targeting MUC16 which is highly expressed on ovarian tumors with limited normal tissue expression; (ii) membrane bound IL-15 (mbIL15) for enhanced in vivo expansion and persistence; and (iii) a kill switch for improved therapeutic control. Healthy donor T cells were transfected with non-viral system encoding CAR, mbIL15 and kill switch to generate PRGN-3005 cells. Expression of transgenes was confirmed by flow cytometry and western blot analyses. PRGN-3005 demonstrated a robust MUC16-specific cytotoxicity of ovarian cancer cells in vitro. PRGN-3005 exhibited a stem-cell like memory phenotype, significant improvement in survival even in absence of exogenous cytokines and lacked autonomous proliferation in vitro in the absence of MUC16 compared to conventional CAR-T cells, demonstrating beneficial effects of mbIL15 on phenotype and persistence of UltraCAR-T cells. Furthermore, we demonstrated that PRGN-3005 cells were selectively and effectively eliminated by kill switch activator antibody via ADCC. For in vivo anti-tumor efficacy of PRGN-3005, an ovarian cancer model of MUC16+ SK-OV-3 cell line was evaluated in NSG mice. In tumor-bearing mice, a single administration of PRGN-3005 one day after gene transfer resulted rapid in vivo expansion and significant increase in UltraCAR-T cells, in particular T stem cell memory (TSCM) population compared to mbIL15neg CAR-T cells. PRGN-3005 demonstrated significantly higher anti-tumor effect (p<0.01) compared to mbIL15neg CAR-T and led to complete tumor regression in all treated mice. PRGN-3005 UltraCAR-T administered one day after non-viral gene transfer has superior anti-tumor efficacy and persistence compared traditional CAR-T cells and represent a promising opportunity for ovarian cancer treatment. Based on these results, FDA has approved an IND application and the first in human PRGN-3005 Phase I clinical trial for advanced ovarian cancer is currently under evaluation (NCT03907527). Citation Format: Tim Chan, Marion Chakiath, Lindsey Shepard, Simon Metenou, Fernando Carvajal-Borda, Jose Velez, Adeline Govekung, Jacques Plummer, Carol Poortman, Padma Kamineny, Xiaohong Ma, Rutul R. Shah, Mary L. Disis, Helen Sabzevari. PRGN-3005 UltraCAR-T™: multigenic CAR-T cells generated using non-viral gene delivery and rapid manufacturing process for the treatment of ovarian cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6593.

Abstract 2180: Genetically engineered chimeric antigen receptor (CAR) monocytes demonstrate targeted anti-tumor activity and differentiate into M1-polarized CAR macrophages

Abstract Despite rapid advances in cancer immunotherapy, clinical responses in metastatic solid tumors have been limited. Macrophages are the most abundant immune cell in the solid tumor microenvironment (TME) and are primarily recruited as monocytes by TME-derived chemokines. When not under the control of the TME, macrophages are potent immune effector cells capable of phagocytosis, T cell recruitment, and antigen presentation. We have previously demonstrated that CAR macrophages (CAR-M) have potent anti-tumor activity and overcome several of the barriers to success in solid tumor immunotherapy - trafficking, immunosuppression, and antigen heterogeneity. Currently, CAR-M are generated via ex vivo differentiation of peripheral blood monocytes into macrophages prior to genetic manipulation. To more closely recapitulate normal biologic behavior, we attempted to create CAR monocytes that could traffic and differentiate into CAR macrophages upon tumor penetration. Toward that goal, we genetically engineered CD14+ human monocytes without ex vivo differentiation and with minimal cell culture. Using the chimeric adenoviral vector Ad5f35, we engineered human CAR-monocytes targeted against HER2. CAR expression and viability both exceeded 90%. Ad5f35 transduced CAR monocytes survived and maintained CAR expression ex vivo for at least 21 days. CAR monocytes efficiently differentiated into CAR-expressing macrophages when treated with GM-CSF as determined by FACS-based phenotypic characterization and Wright-Giemsa staining. Anti-HER2 CAR monocytes eradicated HER2 expressing tumor cells in a time and dose-dependent manner, and had comparable potency to anti-HER2 CAR-M. Additionally, the CAR monocyte manufacturing process offered the logistical advantage of a short manufacturing process (approximately two days). We have previously demonstrated that CAR-M are polarized toward a pro-inflammatory M1 phenotype after transduction with Ad5f35. Similarly, CAR monocytes demonstrated elevated expression of M1 markers, and intriguingly after differentiation into CAR-expressing macrophages, HLA-DR, CD80, CD86, and other M1 markers remained elevated - suggesting that transduction prior to differentiation does not impact the pro-inflammatory impact of adenoviral vectors on myeloid cells. Taken together, this abstract describes the successful development of CAR-monocytes with the potential for a rapid manufacturing process. In addition to direct anti-tumor activity while in the monocyte phase, CAR monocytes have the capacity to differentiate into CAR macrophages in situ, which are in turn capable of phagocytosis, T cell recruitment, TME activation, and antigen presentation. Given the previously demonstrated pre-clinical efficacy of CT-0508 (an anti-HER2 CAR macrophage), the CAR monocyte platform described herein offers a shortened manufacturing process and a potential advantage in tumor penetration, which will be directly evaluated in upcoming studies. Citation Format: Konrad Gabrusiewicz, Maggie Schmierer, Andrew Best, Martha Zeeman, Yumi Ohtani, Linara Gabitova, Daniel Cushing, Saar Gill, Michael Klichinsky. Genetically engineered chimeric antigen receptor (CAR) monocytes demonstrate targeted anti-tumor activity and differentiate into M1-polarized CAR macrophages [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2180.

Effect of Process Parameters on Formability of a AZ31 Magnesium Alloy Thin-Walled Cylindrical Part Formed by Multistage Warm Single-Point Incremental Forming

Magnesium alloys have many distinguished advantages, therefore, they are more and more popularly used in lightweight design of automotive and aviation manufacturing industry. However, its poor plasticity at room temperature has prevented its further application, especially in magnesium alloy sheets forming process. To expand the application of magnesium alloy sheets, single point incremental forming process for rapid prototype manufacturing and small scale productions of sheet metal was investigated. Combining finite element numerical simulations with physical experiments, the relationships between the maximum thickness differences and different process parameters are explored and the optimal process parameters for forming a certain straight wall cylindrical part of AZ31 magnesium alloy were determined. Based on the analysis of the results, the formability of AZ31 magnesium alloy sheet using warm SPIF is improved with the increases of number of forming stage, forming temperature and tool diameter but reduced with the increase of feed rate and interlayer spacing. The suitable forming temperature for AZ31 magnesium alloy sheet is about 250℃. For forming the deeper straight wall cylindrical parts, at least four forming stages are needed.

Solid-State Sensor Chip Produced with Single Laser Engraving for Urine Acidity and Total Dissolved Ion Detections

This paper presents a solid-state sensor for rapidly detecting the acidity and the total dissolved ions of urine. The sensor is produced by direct laser engraving the commercial touch panel film to pattern the sensing areas. Since the indium tin oxide on the touch panel film is an n-type degenerate semiconductor, hydrogen ions in the solution spontaneously attach onto the ITO surface and change the zeta potential of it. The sensing area is connected to the gate of a commercial N-MOSFET CD4007 to establish the extended-gate FET sensing scheme. Full ITO-based sensing chips for pH and total dissolved ions in urine can be easily produced in the developed rapid manufacturing process. Home-build circuits embedded in hand-held devices are produced for the measurement. Results show that the developed ITO-based sensor exhibits high sensing performance for acidity measurement of 0.11 V pH−1 with rapid responses of less than 1 s and low temperature drifting of 0.0205 pH °C−1. The sensor can also detect total dissolved ions with a concentration of 0.1−30 mS in urine. The performance of the developed sensor is characterized by a classic physiology experiment of kidney elution after taking various liquids of reverse osmosis water, beer, and normal saline.

Integrated 3D bioprinting-based geometry-control strategy for fabricating corneal substitutes.

The shortage of donor corneas is a severe global issue, and hence the development of corneal alternatives is imperative and urgent. Although attempts to produce artificial cornea substitutes by tissue engineering have made some positive progress, many problems remain that hamper their clinical application worldwide. For example, the curvature of tissue-engineered cornea substitutes cannot be designed to fit the bulbus oculi of patients. To overcome these limitations, in this paper, we present a novel integrated three-dimensional (3D) bioprinting-based cornea substitute fabrication strategy to realize design, customized fabrication, and evaluation of multi-layer hollow structures with complicated surfaces. The key rationale for this method is to combine digital light processing (DLP) and extrusion bioprinting into an integrated 3D cornea bioprinting system. A designable and personalized corneal substitute was designed based on mathematical modelling and a computer tomography scan of a natural cornea. The printed corneal substitute was evaluated based on biomechanical analysis, weight, structural integrity, and fit. The results revealed that the fabrication of high water content and highly transparent curved films with geometric features designed according to the natural human cornea can be achieved using a rapid, simple, and low-cost manufacturing process with a high repetition rate and quality. This study demonstrated the feasibility of customized design, analysis, and fabrication of a corneal substitute. The programmability of this method opens up the possibility of producing substitutes for other cornea-like shell structures with different scale and geometry features, such as the glomerulus, atrium, and oophoron.

Supply–Demand Prediction for Agile Manufacturing with Deep Neural Network

Abstract Agile manufacturing represents modern production systems that rapidly respond to a fast-moving marketplace and connect customers to production. The success of an agile manufacturing system relies on multisource data analytics, which provide smart data-driven decision-making strategies to guide manufacturing and the supply chain system. In order to implement rapid manufacturing processes to respond to customer orders, supply–demand gap prediction plays a critical role. In this article, we study the problem of predicting supply–demand gap with respect to product types, categories, and spatiotemporal attributes. To this end, we propose and develop an end-to-end framework using an extendable deep neural network architecture for supply–demand gap reduction. The framework can process multiple customized input factors and automatically discover spatiotemporal supply–demand patterns from historical transaction data. A set of customized features are employed to build a general training model to predict future demand. Embedding layers are used to map high dimensional features into a smaller subspace for input data consolidation. Fully connected layers with activation functions are used to build the training architecture of the model. Customized data attributes can be concatenated from different layers in the deep learning neural network. In this way, multisource input data can be integrated with outputs of internal layers for a comprehensive demand prediction. Experiments were conducted to evaluate the network with real supply and demand data, which were acquired from warehouses of a manufacturing company. The experimental results demonstrated that the proposed network was able to reduce supply–demand gap and provide a practical solution to long-term customer demand prediction.

ARIMA-GMDH: a low-order integrated approach for predicting and optimizing the additive manufacturing process parameters

This paper proposes a novel data-driven approach for predicting and optimizing the additive manufacturing process parameters. The integrated scheme consists of three popular algorithms: (1) group method for data handling (GMDH) as the engine of neural networks, (2) autoregressive integrated moving average (ARIMA) for characterizing spatial collinearity of the multiple response, and (3) indirect optimization on the basis of self-organization (IOSO) to adopt the emerged correlated multi-response optimization problem. As a numerical case study, a computer-generated fused deposition modeling data tested the introduced algorithms. The finite element (FE) simulation model consists the multi-layer residual stresses as targets, in respect of printing speeds as process parameters. The residual stresses predicted by the low-order integrated ARIMA-GMDH variants correlate well with the FE simulations. This approach provides a viable data-driven alternative for computationally based rapid prototyping and additive manufacturing processes.

Preclinical Characterization of Prgn-3006 Ultracar-T™ for the Treatment of AML and MDS: Non-Viral, Multigenic Autologous CAR-T Cells Administered One Day after Gene Transfer

Relapsed/refractory (r/r) acute myeloid leukemia (AML) is an aggressive malignancy with poor prognosis and limited treatment options underscoring the need for new therapies. Traditional methods for generating CAR-T cells require extensive ex vivo expansion following viral vector transduction, adding time and high cost to manufacturing with a centralized manufacturing process and prolonging the waiting period between apheresis and administration of CAR-T therapy to a patient. Time is of the essence for advanced cancer patients including patients with r/r AML. The UltraCAR-T™ platform addresses shortcomings of current CAR-T technologies by streamlined rapid manufacturing process that can be performed in a local hospital's GMP facility following apheresis from the patient providing a potential solution for broader adoption of CAR-T therapies. The UltraCAR-T™ platform utilizes Precigen's advanced non-viral multi-gene delivery system and the rapid manufacturing process with high cell viability that allows for administration of autologous CAR-T cells one day after gene transfer. PRGN-3006 UltraCAR-T™ cells, utilizing Precigen's UltraVector® platform, co-express CD33 CAR, membrane bound IL-15 (mbIL15) and a kill switch. Expression of mbIL15 provides CAR-T cells with a source of cytokine without the need for exogenous cytokines to further enhance the expansion potential in the presence of tumor antigen and the overall persistence for PRGN-3006 in. The expression of mbIL15 on PRGN-3006 eliminates any need for ex vivo expansion of T cells prior to cell administration. Co-expression of kill switch provides a mechanism for selective depletion of PRGN-3006 post administration and improves therapeutic control. We manufactured PRGN-3006 UltraCAR-T™ cells using non-viral gene transfer and rapid streamlined manufacturing process from multiple donor T cells. Co-expression of CAR, mbIL15 and kill switch was confirmed by flow cytometry and western blot analyses. PRGN-3006 demonstrated robust expansion in presence of CD33 antigen, lack of autonomous expansion in absence of CD33, and prolonged persistence in absence of exogenous cytokines in in vitro tests. The redirected specificity was demonstrated by the specific killing of CD33+ tumor cells as well as significant release of inflammatory cytokines such as IFNγ upon co-culture with AML tumor cells. We also demonstrated specific elimination of PRGN-3006 cells by kill switch activator antibody treatment. In vivo, a single administration of PRGN-3006 UltraCAR-T™ cells only one day after gene transfer effectively eliminated tumor burden and significantly improved overall survival (p<0.01) of tumor bearing mice compared to CAR-T cells lacking mbIL15 expression (conventional CAR-T) in an aggressive xenograft model of AML in immunocompromised (NSG) mice. PRGN-3006 demonstrated engraftment and significantly higher expansion and persistence in mice compared to CAR-T cells lacking mbIL15 (conventional CAR-T). These pre-clinical data provide a strong rationale to evaluate PRGN-3006 UltraCAR-T™ in a clinical trial for treatment of AML. The FDA has approved an investigational new drug application for PRGN-3006 UltraCAR-T™ and the first in human Phase 1 clinical trial for treatment of patients with relapse/refractory AML and high-risk myeloid dysplastic syndrome is currently ongoing (ClinicalTrials.gov Identifier: NCT03927261). Figure: Antitumor activity of PRGN-3006 UltraCAR-T™ cells in an established model of AML. MOLM-13 tumor cells were engrafted into NSG mice on Day 0 and PRGN-3006 were administered on Day 6 (arrow). Data shown is tumor burden (expressed as total flux in photons/sec) as the mean±SEM for each group of mice shown, with n=6-7 mice mice/group at the start of the study. Figure Disclosures Chan: Precigen: Employment. Ma:Precigen, Inc.: Employment. Carvajal-Borda:Precigen, Inc.: Employment. Velez:Precigen, Inc.: Employment. Plummer:Precigen, Inc.: Employment. Shepard:Precigen, Inc.: Employment. Shah:Precigen, Inc.: Employment; Intrexon Corp.: Equity Ownership. Sallman:Celyad: Membership on an entity's Board of Directors or advisory committees. Sabzevari:Precigen, Inc.: Employment.