Manufacturing Challenges Research Articles

Development of Q&P steels of third generation under conditions simulating a continuous annealing process, and evaluation of their weldability by the RSW process

Quenching and partitioning, advanced high-strength steels (Q&P-AHSS), are considered as potential candidates to satisfy the current requirements of the automotive industry in terms of passenger safety. Their fabrication at industrial scale has required modifications to well-established processes and thus, obtaining this type of microstructures through easier thermal cycles represents a current challenge for steel manufacturers. Apart from their manufacturing method, their mechanical properties and weldability are equally important. In the present work, Q&P-AHSS were obtained through short-term annealing and isothermal treatments, to further investigate the effects of resistance spot welding (RSW) parameters, on nugget quality and tensile shear strength. Results show that Q&P-AHSS can be welded with success by the RSW process, meeting the required quality specified in the standard AWS D8.9 M-2012.

Is read-across for chemicals comparable to medical device equivalence and where to use it for conformity assessment?

Novel medical devices must conform to medical device regulation (MDR) for European market entry. Likewise, chemicals must comply with the Registration, Evaluation, Authorization and Restriction of Chemicals (REACh) regulation. Both pose regulatory challenges for manufacturers, but concordantly provide an approach for transferring data from an already registered device or compound to the one undergoing accreditation. This is called equivalence for medical devices and read-across for chemicals.Although read-across is not explicitly prohibited in the process of medical device accreditation, it is usually not performed due to a lack of guidance and acceptance criteria from the authorities. Nonetheless, a scientifically justified read-across of material-based endpoints, as well as toxicological assessment of chemical aspects, such as extractables and leachables, can prevent failure of MDR device equivalence if data is lacking. Further, read-across, if applied correctly can facilitate the standard MDR conformity assessment.The need for read-across within medical device registration should let authorities to reconsider device accreditation and the formulation of respective guidance documents. Acceptance criteria like in the European Chemicals Agency (ECHA) read-across assessment framework (RAAF) are needed. This can reduce the impact of the MDR and help with keeping high European innovation device rate, beneficial for medical device patients.

Full-composition-gradient in-situ alloying of Cu–Ni through laser powder bed fusion

Multi-material laser powder bed fusion has long been a challenge in additive manufacturing, especially in terms of controlling spatial variations and joining multiple materials. The challenge is even greater when multi-material components have different physical properties (e.g., different thermal expansions, thermal conductivities and laser reflectivities). The aforementioned scenario is applicable to copper–nickel (Cu–Ni) alloy. The simultaneous printing and controllable mixing of pure Cu and pure Ni within a single print have not been accomplished due to the substantial disparity in the physical properties of pure Cu and pure Ni. In this work, a full composition with different mixing proportions (100 % Cu to 100 % Ni) was fabricated in a single printing process through in-situ alloying based on our process of micro laser powder bed fusion. A transition from a columnar grain to a quasi-equiaxed grain morphology with texture variation from <110> to <111> was found in the full–composition–gradient of the Cu–Ni alloy. Microstructural variations, such as fine grain strengthening and grain boundary strengthening, greatly affected the mechanical properties of the alloys: the ultimate tensile strength ranged from 303 to 488 MPa and the yield strength ranged from 231 to 445 MPa. Moreover, the electrical conductivity of the alloys with a compositional gradient ranged from 3.6 % IACS to 96 % IACS due to the electron scattering induced by lattice distortion. In summary, a full-composition Cu–Ni gradient alloy was fabricated through in-situ alloying adopting micro laser powder bed fusion for the first time, enabling the investigation of the evolution of the microstructure and properties of the alloy as the alloy composition varies.

Development of an online optical prototype for the simultaneous analysis of defects or gels in industrially synthesized polypropylene films. Part 1: Comparison with ASTM D 3351–93 methods

Polypropylene (PP) films are crucial in various industrial applications, from packaging to medical products. However, a common challenge in PP manufacturing is the presence of gel-like defects. These gels are minor defects on the surface of the films that significantly affect the physicochemical, mechanical, and organoleptic properties of the films, compromising the quality of the final product. This first research focuses on developing and validating an in-line optical method to replace the international method ASTM D 3351–93. The main objective was to create a methodology that has the same scope and analytical performance as those reported by ASTM D 3351–93 in such a way that it can compete with it in terms of precision and accuracy, thus allowing end users to this ASTM, such as PP producers, PP marketers, PP film producers, among others internationally, can use this new methodology with necessary analytical support. This analytical methodology integrates the PP extrusion zones, the film processing stages, and the optical zone for reading and processing analytical data. Additionally, it has the advantage of working with a sample size that is even more representative of the population and has less human error since only one operator is required to carry out the test; this method also has much shorter response times. The developed prototype had 14 online stages that allowed representative quantities of samples to be taken and processed thermally and mechanically for ideal optical measurement. For the online method, a 6-point calibration curve is carried out at concentrations of 40, 10, 5, 2, 1 and 0 ppm for the gel or defect sizes of 200, 400, 500, 600, 700, 800 and 900 µm, showing excellent linearity where the correlation coefficient varied between 0.997 and 0.999, the limits of detection (LOD) varied between 0.85 and 2.61 and the limits of quantification (LOQ) ranged between 2.82 and 8.71. The statistical analyzes by ANOVA of the comparison between the ASTM D 3351–93 method and the proposed simultaneous method indicate that the p value of the evaluation of the means was 0.946, which suggests that the means are not statistically different. To complement, the Tukey test was carried out at 95 %, indicating that the methods have statistical equivalence.•Process optimization•Determination of defects or imperfections in PP films

Chemical management in electronics manufacturing: Protecting worker health and the environment

In the realm of electronics manufacturing, the management of chemicals is paramount to safeguarding both worker health and environmental sustainability. This review delves into the strategies and challenges associated with chemical management within this industry. The utilization of various chemicals in electronics manufacturing processes presents potential hazards to both workers and the environment. From cleaning agents to solvents and fluxes, these substances pose risks ranging from acute toxicity to long-term health effects and environmental contamination. Effective chemical management strategies are therefore indispensable. This review discusses the proactive measures implemented by electronics manufacturers to mitigate these risks. It explores the adoption of alternative, less hazardous chemicals and the implementation of engineering controls to minimize exposure. Additionally, stringent protocols for handling, storage, and disposal are essential components of comprehensive chemical management programs. Furthermore, regulatory frameworks play a pivotal role in shaping chemical management practices within the electronics manufacturing sector. Compliance with local and international regulations such as REACH (Registration, Evaluation, Authorization, and Restriction of Chemicals) and RoHS (Restriction of Hazardous Substances) is imperative to ensure the safety of workers and the environment. However, despite these efforts, challenges persist. Balancing the need for innovation and productivity with chemical safety requirements remains a significant challenge for manufacturers. Additionally, global supply chain complexities add another layer of complexity to chemical management efforts. Effective chemical management in electronics manufacturing demands a multi-faceted approach encompassing technological innovation, regulatory compliance, and a commitment to worker health and environmental stewardship. By addressing these challenges collaboratively, the industry can strive towards safer and more sustainable practices.

A Model Study to Assess Fibrillation and Product Stability to Support Peptide Drug Design.

The fibrillation of therapeutic peptides can present significant quality concerns and poses challenges for manufacturing and storage. A fundamental understanding of the mechanisms of fibrillation is critical for the rational design of fibrillation-resistant peptide drugs and can accelerate product development by guiding the selection of solution-stable candidates and formulations. The studies reported here investigated the effects of structural modifications on the fibrillation of a 29-residue peptide (PepA) and two sequence modified variants (PepB, PepC). The C-terminus of PepA was amidated, whereas both PepB and PepC retained the carboxylate, and Ser16 in PepA and PepB was substituted with a helix-stabilizing residue, α-aminoisobutyric acid (Aib), in PepC. In thermal denaturation studies by far-UV CD spectroscopy and fibrillation kinetic studies by fluorescence and turbidity measurements, PepA and PepB showed heat-induced conformational changes and were found to form fibrils, whereas PepC did not fibrillate and showed only minor changes in the CD signal. Pulsed hydrogen-deuterium exchange mass spectrometry (HDX-MS) showed a high degree of protection from HD exchange in mature PepA fibrils and its proteolytic fragments, indicating that most of the sequence had been incorporated into the fibril structure and occurred nearly simultaneously throughout the sequence. The effects of the net peptide charge and formulation pH on fibrillation kinetics were investigated. In real-time stability studies of two formulations of PepA at pH's 7.4 and 8.0, analytical methods detected significant changes in the stability of the formulations at different time points during the study, which were not observed during accelerated studies. Additionally, PepA samples were withdrawn from real-time stability and subjected to additional stress (40 °C, continuous shaking) to induce fibrillation; an approach that successfully amplified oligomers or prefibrillar species previously undetected in a thioflavin T assay. Taken together, these studies present an approach to differentiate and characterize fibrillation risk in structurally related peptides under accelerated and real-time conditions, providing a model for rapid, iterative structural design to optimize the stability of therapeutic peptides.

Adeno-associated virus genome quantification with amplification-free CRISPR-Cas12a.

Efficient manufacturing of recombinant Adeno-Associated Viral (rAAV) vectors to meet rising clinical demand remains a major hurdle. One of the most significant challenges is the generation of large amounts of empty capsids without the therapeutic genome. There is no standardized analytical method to accurately quantify the viral genes, and subsequently the empty-to-full ratio, making the manufacturing challenges even more complex. We propose the use of CRISPR diagnostics (CRISPR-Dx) as a robust and rapid approach to determine AAV genome titers. We designed and developed the CRISPR-AAV Evaluation (CRAAVE) assay to maximize sensitivity, minimize time-to-result, and provide a potentially universal design for quantifying multiple transgene constructs encapsidated within different AAV serotypes. We also demonstrate an on-chip CRAAVE assay with lyophilized reagents to minimize end user assay input. The CRAAVE assay was able to detect AAV titers as low as 7e7 vg/mL with high precision (<3% error) in quantifying unknown AAV titers when compared with conventional quantitative PCR (qPCR) method. The assay only requires 30 min of assay time, shortening the analytical workflow drastically. Our results suggest CRISPR-Dx could be a promising tool for efficient rAAV genome titer quantification and has the potential to revolutionize biomanufacturing process analytical technology (PAT).

Machining technologies and structural models of microfluidic devices

In the past decades, microfluidic chips have been one of the hottest research topics in the “lab-on-a-chip” field. However, it is still a challenge for design the perfect microchannels with high functional integration, complex geometry, and high production efficiency. This review paper provides a comprehensive overview of microfluidic structural models and designs, manufacturing techniques, and bonding methods. The effect of three types of microfluidic channel structure models and their configuration parameters on the fluid behaviors of microfluidics was investigated. Surface-to-volume ratio of the flow channel is a critical parameter for determining the mixing and diffusion capabilities of the microfluidic devices (MDs). Thereafter, the main manufacturing techniques for microstructures were discussed, including etching, photolithography, soft lithography, molding, mechanical micro-machining, laser beam micromachining, printing, and 3D printing. We performed a statistical analysis to discuss the relative advantages and disadvantages of these manufacturing methods. In addition, the current bonding methods for MDs were also summarized, including thermal bonding, solution adhesive bonding, dry adhesive bonding, plasma bonding, and anodic bonding. a comparison of different bonding methods was presented to provide a reference to select a suitable bonding method for the assembly of microfluidic device. Finally, the tendencies and challenges of microchannel manufacturing were discussed, and the prospects were also provided.

Abstract 2714: Developing an mRNA encoded therapeutic antibody platform to simplify manufacturing and reduce time to clinic

Abstract The manufacturing of therapeutic antibodies requires expensive production, often challenging purification, lengthy stability optimization and complex protein characterization that despite continual improvement, keeps the cost of treatment in the clinic high. Alternatively, leveraging gene-based approaches such as mRNA to express therapeutics circumvents many of the manufacturing challenges and instead rely on in situ production of antibodies within a patient, improving the developability and cost of sophisticated, disease-modifying antibodies. Crucially, recent work demonstrates that therapeutic antibodies translated in vivo from mRNA can be readily detected within hours following infusion into pre-clinical models and can persist up to several days or weeks. Peak levels of circulating mRNA-encoded antibodies are comparable to infused recombinant equivalents dosed to patients and have been shown to be within favorable therapeutic ranges in phase I trials. Here we outline an mRNA-LNP based platform to encode and deliver therapeutic antibodies in vivo that can overcome costly and challenging manufacturing and that demonstrates both robust PK/PD kinetics and potency. We describe how to effectively encode a standard-of-care anti-HER2 antibody - Trastuzumab - using LNP-encapsulated modified mRNA and validate using in vitro as well as in vivo efficacy models. Initial in vitro characterization of mRNA encoded antibodies demonstrates robust translation in producer cell lines, reproducible expression kinetics, retention of antigen specificity and HC:LC integrity when analyzed by ELISA and WB. To evaluate whether Trastuzumab-encoding mRNA-produced protein is detectable in vivo, antibody levels were measured in mouse serum after IV administration of a single dose of mRNA formulated LNPs. Peak plasma concentrations of mRNA-encoded trastuzumab were benchmarked against circulating recombinant Trastuzumab in parallel groups, and pharmacokinetics interrogated. Of note, the majority of mRNA encoded antibody was likely produced in the liver as shown by bioluminescence imaging. To demonstrate the efficacy of the platform, the potency of mRNA-encoded Trastuzumab isolated from producer cell lines will be assessed using in vitro cytotoxicity assays with human PBMCs and HER2+ tumor lines to dissect ADCC or direct tumor growth inhibition. Additionally, to determine whether mRNA-encoded Trastuzumab is able to retain anti-tumor activity in vivo, a mouse tumor xenograft model will be established and the mRNA antibody-dependent effects on tumor volume, growth and morbidity-free survival will be examined. Overall, the data demonstrate that mRNA-encoded therapeutic antibodies could provide an effective, alternate strategy for solid tumor treatment and may unlock a strategy to deliver lead biologics at reduced cost and with improved developability. Citation Format: Roxana Redis, Dan Rocca, Rachel Pooley, Matthew Benson, Namrata Jayanth, Eva Oswald, Alexander Hale, Louise Brackenbury, Justin Bryans. Developing an mRNA encoded therapeutic antibody platform to simplify manufacturing and reduce time to clinic [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2714.

A fast simulation method for thermal management in wire arc additive manufacturing repair of a thin-walled structure

Ensuring first-time-right on-site repair of critical structures is a key challenge for additive manufacturing (AM)–based repair solutions. Fast thermal simulations are thus needed to plan efficient and error-free AM processes. This paper addresses a fast thermal simulation method for a novel subsea wire arc additive manufacturing (SWAAM) repair procedure. Current commercial finite element (FE) codes for typical welding and AM are computationally expensive and slow. The presented 2D finite difference approach can be used to simulate SWAAM on a damaged plate with around 70 times acceleration compared to real welding times, without the use of parallelization. Although not being able to accurately represent the temperature in close vicinity of the welding torch, the approach shows excellent correspondence with FE simulations and experiments in regions of the plate where the temperature has assumed a distribution that is largely two-dimensional. Compared with FE simulations, the approach is experimentally verified to be accurate to 10 °C within 7 s after the welding torch has passed a point on the plate. Thus, the approach can provide a measure of the global temperature field in a thin-walled structure during repair. The thermal simulation is preceded by a welding path planner, which generates appropriate paths based on slicing of a 3D surface scan of the damage that is to be repaired. Damages to equipment or non-ideal welding conditions are prevented by automatically pausing the welding if the calculated temperature in the path ahead of the welding torch exceeds a predefined interpass temperature limit.

Formulation of inks for 3D printing of microalgae‐based meat analogues and the role of modified starch: a review

SummaryThe quest for sustainable alternatives to traditional meat products has led to increased interest in plant‐based meat analogues. Microalgae, with their high protein content and eco‐friendly cultivation methods, present a promising option for the development of such analogues. However, integrating microalgae into meat analogues poses challenges related to achieving desired texture, flavour, and colour. This review explores the potential of modified starch (MS) as a solution to enhance the quality of microalgae‐based meat analogues produced via 3D printing. Key findings from the manuscript indicate that MSs can improve the viscosity, elasticity, and printability of microalgae‐based edible inks. Furthermore, they play a crucial role in reducing off‐flavours and off‐colours in the final product. By optimising the type and concentration of MSs used, microalgae‐based meat analogues can be tailored to meet consumer preferences while addressing environmental concerns. Overall, the utilisation of MSs represents a practical approach to overcoming manufacturing challenges associated with microalgal‐based meat analogues, paving the way for the development of sustainable and nutritious plant‐based meat products through innovative and ecologically friendly food technologies.

Recent progress of polymeric microneedle-assisted long-acting transdermal drug delivery.

Microneedle (MN)-assisted drug delivery technology has gained increasing attention over the past two decades. Its advantages of self-management and being minimally invasive could allow this technology to be an alternative to hypodermic needles. MNs can penetrate the stratum corneum and deliver active ingredients to the body through the dermal tissue in a controlled and sustained release. Long-acting polymeric MNs can reduce administration frequency to improve patient compliance and therapeutic outcomes, especially in the management of chronic diseases. In addition, long-acting MNs could avoid gastrointestinal reactions and reduce side effects, which has potential value for clinical application. In this paper, advances in design strategies and applications of long-acting polymeric MNs are reviewed. We also discuss the challenges in scale manufacture and regulations of polymeric MN systems. These two aspects will accelerate the effective clinical translation of MN products.

Revolutionizing plasmonic platform via magnetic field-assisted confined ultrafast laser deposition of high-density, uniform, and ultrafine nanoparticle arrays

The remarkable capabilities of 2D plasmonic surfaces in controlling optical waves have garnered significant attention. However, the challenge of large-scale manufacturing of uniform, well-aligned, and tunable plasmonic surfaces has hindered their industrialization. To address this, we present a groundbreaking tunable plasmonic platform design achieved through magnetic field (MF) assisted ultrafast laser direct deposition in air. Through precise control of metal nanoparticles (NPs), with cobalt (Co) serving as the model material, employing an MF, and fine-tuning ultrafast laser parameters, we have effectively converted coarse and non-uniform NPs into densely packed, uniform, and ultrafine NPs (∼3 nm). This revolutionary advancement results in the creation of customizable plasmonic ‘hot spots,’ which play a pivotal role in surface-enhanced Raman spectroscopy (SERS) sensors. The profound impact of this designable plasmonic platform lies in its close association with plasmonic resonance and energy enhancement. When the plasmonic nanostructures resonate with incident light, they generate intense local electromagnetic fields, thus vastly increasing the Raman scattering signal. This enhancement leads to an outstanding 2–18 fold boost in SERS performance and unparalleled sensing sensitivity down to 10−10 M. Notably, the plasmonic platform also demonstrates robustness, retaining its sensing capability even after undergoing 50 cycles of rinsing and re-loading of chemicals. Moreover, this work adheres to green manufacturing standards, making it an efficient and environmentally friendly method for customizing plasmonic ‘hot spots’ in SERS devices. Our study not only achieves the formation of high-density, uniform, and ultrafine NP arrays on a tunable plasmonic platform but also showcases the profound relation between plasmonic resonance and energy enhancement. The outstanding results observed in SERS sensors further emphasize the immense potential of this technology for energy-related applications, including photocatalysis, photovoltaics, and clean water, propelling us closer to a sustainable and cleaner future.

Development, Prospect and Challenge for Additive Manufacturing of Polymer Materials

As a manufacturing technology that fabricates three-dimensional objects by stacking materials layer by layer, additive manufacturing can be divided into laser fused deposition, electron beam fused deposition, inkjet printing and so on based on different additive manufacturing processes. In recent years, remarkable progress has been made in the additive manufacture of polymer materials. Firstly, the additive manufacturing equipment and process parameters suitable for polymer materials have been successfully developed, which realized the accurate manufacturing of complex structures. Secondly, by optimizing the formula and post-treatment of polymer materials, the performance and durability of additive products are improved. In addition, the researchers also explored the application potential of additive manufacturing of polymer materials in medical, aerospace and other fields. However, the additive manufacturing of polymer materials still faces some challenges. One of them is that the rheological properties of polymer materials have a great impact on the progress of additive manufacturing, which needs further research and improvement. Meanwhile, the heat and stress generated in additive manufacturing may lead to the deformation and degradation of polymer materials, which requires effective thermal management. Finally, compared with the traditional cutting process, the additive manufacturing cost of polymer materials is still high, so it is necessary to reduce the cost and promote production efficiency. In a word, additive manufacturing of polymer materials has broad prospects and application potential, but further research and improvement are still needed to overcome related challenges.

MitM attacks on intellectual property and integrity of additive manufacturing systems: A security analysis

Additive Manufacturing (AM) was originally invented to reduce the cost of the prototyping process. Over time, the technology evolved to be faster, more accurate, and affordable. These factors, in addition to the potential use of AM in parts production, have helped rapidly drive the growth of AM in both industrial and personal uses. Thus, there is an accompanying demand to understand the cybersecurity implications of such systems. In our research, we present an in-depth security review of Stratasys Dimension Elite and show how manufacturers of such high-end 3D printers failed to protect the confidentiality and integrity of the printed 3D models. Revealing the intricate dimensions of cyber threats within the realm of AM and laying the foundation for understanding the multifaceted nature of attacks, offering insights into vulnerabilities and potential consequences. Moreover, we demonstrate the massive impact network attacks can have on 3D printers' communication channels. Our sniffing attack stole transmitted models with a minimal overhead of 0.015 seconds to evade detection. The developed replacement attack targeted and replaced specific models with offline-prepared models. Also, we automated a sabotaging attack to alter the interior model structure on the fly with minimal visual but significant strength differences. By revealing these attacks, this research not only improves the security posture of 3D printers but also enhances the understanding of security challenges in additive manufacturing as a whole.

From inspection to analysis: A combined approach to identifying counterfeit whiskeys using HS-GC-FID and bottle integrity

Counterfeiting of alcoholic beverages, particularly high-value spirits such as whiskey, presents significant challenges for regulators, manufacturers, and consumers. In this study, we introduce and validate a novel application of headspace extraction (HS) followed by gas chromatography with flame ionization detection (GC-FID) for the quantitative determination of ethanol content in 42 suspected counterfeit brazilian samples of whiskeys. This method, in conjunction with visual inspection of material inconsistencies, offers a combined approach to identify potential cases of fraud. The HS-GC-FID findings revealed that only 19% of the analyzed samples had ethanol content in the limits declared on the label, emphasizing the role of ethanol content as a chemical marker for suspected beverage fraud.

A self-supervised learning framework based on masked autoencoder for complex wafer bin map classification

Wafer bin map (WBM) automatic classification is one of the critical challenges for semiconductor intelligent manufacturing. Many deep learning-based classification models have performed well in WBM classification, but all require a large amount of labeled data for training. Since real-world WBMs are highly complex and can be labeled correctly only by seasoned engineers, such requirements undermine the practical value of those methods. Several self-supervised learning methods have recently been proposed for WBM to improve classification performance. However, they still require much labeled data for fine-tuning and are only adapted for binary WBM with a single gross failure area. To address these limitations, this study introduces a self-supervised framework based on masked autoencoder (MAE) for complex WBMs with mixed bin signatures and multiple gross failure area patterns. A patchMC encoder is proposed to improve MAE’s representation ability for complex WBMs with mixed bin signatures. Moreover, the pre-trained MAE encoder with a multi-label classifier fine-tuned by labeled WBMs enables a few-shot classification of complex WBMs with multiple gross failure areas. Experimental validation of the proposed method is performed on a real-world complex WBM dataset from Intel Corporation. The results demonstrate that the proposed method can make good use of unlabeled WBMs and reduce the demand for labeled data to a few-shot level and, at the same time, guarantees a classification accuracy of more than 90%. By comparing MAE with other self-supervised learning methods, MAE outperforms other existing self-supervised methods for WBM data.

Technological challenges in manufacturing of vacuum gauge thermionic cathode using thick-film technology

Abstract This paper focuses on the development of a technological challenges of manufacturing the planar ceramic vacuum sensor based on the principles of hot-cathode ionization in the Bayard-Alpert configuration. The goal is to simplify the technological process by utilizing planar platinum structures with gold electrical paths instead of 3-dimensional structures. Various methods were tested, including the use of carbon-based SVM (Sacrifice Volume Materials) materials, but without success. Wet-etching using potassium hydroxide on Al2O3 substrates showed promise results. The findings highlight the challenges and progress made in developing the thermo-emittercomponent of the vacuum sensor.

Starting from an Empty Plate: A Semiotic Study of Instant Noodle Advertisements during the Fasting Month of Ramadan in the Covid-19 Pandemic

From the beginning of the Covid-19 pandemic in 2020 until 2022, Indomie has consistently issued advertisements during the fasting month of Ramadan. During this period, it is a challenge for food manufacturers to promote their products. In the case of Indomie’s advertisement, which started with an idea that visually displays an empty plate and is broadcast during the fasting month, is quite viral because it dares to advertise without displaying the products it sells. Interestingly, Indomie is still holding up empty plates for their advertisement in 2021. The continuity of the story that started with empty plates is what prompted me to continue the analysis of Indomie instant noodle ads that were aired during the fasting months of Ramadan during the pandemic in 2020, 2021, and 2022. The purpose of this research is to reveal the meaning and ideology of what is to be conveyed through these ads. Rolland Barthes' semiotic analysis method is considered suitable to analyze the mythology and ideology behind them. The presence of an empty plate shows the meaning of tolerance for fasting Muslims. Each advertisement describes this form of tolerance differently. Even though it is covered by religious, social, cultural, and economic discourses, the ideology of capitalism is still sensed in all the advertisements that are displayed.

Tuning capsid formation dynamics in recombinant adeno-associated virus producing synthetic cell lines to enhance full particle productivity.

Recombinant adeno-associated virus (rAAV) is widely used as an in vivo delivery vector for gene therapy. It is used in a very large dose, and the large quantities required for broad applications present manufacturing challenges. We have developed a synthetic biology platform of constructing cell lines integrated with essential viral genes which can be induced to produce rAAV without plasmid transfection or virus transduction. Through iterative design-construct-characterization cycles, we have showcased the potential of this synthetic cell production system. Systems characterization of the dynamics of viral transcripts and proteins as well as virus assembly and packaging revealed that the expression level and balance of viral genome and capsid protein are keys to not only the productivity but also the full particle content, an important product quality attribute. Boosting cap gene expression by sequential transfection and integration of multiple copies of the cap gene elevated the rAAV titer to levels on a par with traditional plasmid transfection and virus infection. However, overexpression of the cap gene shifted the balance and kinetics of the genome and capsid. We independently tuned the dynamics of genome amplification and capsid protein synthesis by modulating the induction concentration as well as the time profile, and significantly enhanced full particle content while maintaining a high productivity. This strategy of constructing an inducible stable producer cell line is readily adaptable to rAAV vectors of different serotypes and payloads. It can greatly facilitate scalable production of gene therapy vectors.