Stages Of Development Process Research Articles

Use of focused acoustics for cell disruption to provide ultra scale‐down insights of microbial homogenization and its bioprocess impact—recovery of antibody fragments from rec E. coli

An ultra scale-down (USD) device that provides insight of how industrial homogenization impacts bioprocess performance is desirable in the biopharmaceutical industry, especially at the early stage of process development where only a small quantity of material is available. In this work, we assess the effectiveness of focused acoustics as the basis of an USD cell disruption method to mimic and study high-pressure, step-wise homogenization of rec Escherichia coli cells for the recovery of an intracellular protein, antibody fragment (Fab'). The release of both Fab' and of overall protein follows first-order reaction kinetics with respect to time of exposure to focused acoustics. The rate constant is directly proportional to applied electrical power input per unit volume. For nearly total protein or Fab' release (>99%), the key physical properties of the disruptate produced by focused acoustics, such as cell debris particle size distribution and apparent viscosity show good agreement with those for homogenates produced by high-pressure homogenization operated to give the same fractional release. The only key difference is observed for partial disruption of cells where focused acoustics yields a disruptate of lower viscosity than homogenization, evidently due to a greater extent of polynucleic acids degradation. Verification of this USD approach to cell disruption by high-pressure homogenization is achieved using USD centrifugation to demonstrate the same sedimentation characteristics of disruptates prepared using both the scaled-down focused acoustic and the pilot-scale homogenization methods for the same fraction of protein release.

Prediction and verification of centrifugal dewatering of P. pastoris fermentation cultures using an ultra scale‐down approach

Recent years have seen a dramatic rise in fermentation broth cell densities and a shift to extracellular product expression in microbial cells. As a result, dewatering characteristics during cell separation is of importance, as any liquor trapped in the sediment results in loss of product, and thus a decrease in product recovery. In this study, an ultra scale-down (USD) approach was developed to enable the rapid assessment of dewatering performance of pilot-scale centrifuges with intermittent solids discharge. The results were then verified at scale for two types of pilot-scale centrifuges: a tubular bowl equipment and a disk-stack centrifuge. Initial experiments showed that employing a laboratory-scale centrifugal mimic based on using a comparable feed concentration to that of the pilot-scale centrifuge, does not successfully predict the dewatering performance at scale (P-value <0.05). However, successful prediction of dewatering levels was achieved using the USD method (P-value ≥0.05), based on using a feed concentration at small-scale that mimicked the same height of solids as that in the pilot-scale centrifuge. Initial experiments used Baker's yeast feed suspensions followed by fresh Pichia pastoris fermentation cultures. This work presents a simple and novel USD approach to predict dewatering levels in two types of pilot-scale centrifuges using small quantities of feedstock (<50 mL). It is a useful tool to determine optimal conditions under which the pilot-scale centrifuge needs to be operated, reducing the need for repeated pilot-scale runs during early stages of process development.

Conference Scene: Recent in microscale bioprocess

Advances in genetics, as well as molecular and cellular biology, are continually fueling innovations in the biopharmaceutical sector. Increasingly specialized therapies are emerging that offer exciting prospects for the treatment of human diseases. Defining a suitable manufacturing route for each therapeutic product, however, can often present a significant challenge. There is a growing trend in the biopharmaceutical sector to use microscale models (that mimic key manufacturing steps) during the early stages of process development, as such technologies enable key process information to be accrued rapidly and at relatively low costs. The 2nd Annual Miniaturisation conference, organized by Euroscicon, brought together 34 delegates from industry and academia for a lively discussion of the current state-of-the-art in microscale bioprocess technologies.

Sustainability assessment of novel chemical processes at early stage: application to biobased processes

Chemical conversions have been a cornerstone of industrial revolution and societal progress. Continuing this progress in a resource constrained world poses a critical challenge which demands the development of innovative chemical processes to meet our energy and material needs in a sustainable way. This challenge forms the basis for this article. We report a method for quick preliminary assessment of chemical processes at the laboratory stage. The proposed method enables a review of chemical processes within a broader sustainability context. It is inspired by green chemistry principles, techno-economic analysis and some elements of environmental life-cycle assessment (LCA). This method evaluates a proposed chemical process against comparable existing processes using a multi-criteria approach that integrates various economic and environmental indicators. An effort has been made to incorporate quantitative and qualitative information about the processes while making the method transparent and easy to implement based on information available at an early stage in process development. The idea is to provide a data-based assessment tool for chemists and engineers to develop sustainable chemistry. This paper describes the method in detail and examines plausibility of the results. A biobased process for the production of but-1,3-diene has been analyzed using this method. This biobased process is compared with a conventional process for the production of but-1,3-diene from petroleum sources. The effects of uncertainty in the underlying model parameters and assumptions are also analyzed, along with the effect of system boundary selection on the assessment outcome. Analysis and testing of the method shows that it can be used as a valuable tool for sustainable process development.

How to Model a Secure Information System (IS): A Case Study

 Abstract—The existing information system (IS) developments methods are not met the requirements to resolve the security related (IS) problems and they fail to provide a successful integration of security and systems engineering during all development process stages. Hence, the security should be considered during the whole software development process and identified with the requirements specification. This paper aims to propose an integrated security and IS engineering approach in all software development process stages by using i* language. This proposed framework categorizes into three separate parts: modelling business environment part, modelling information technology system part and modelling IS security part. Booking hotel room management process is used as a case study to validate the proposed framework. The results show that considering security IS goals in the whole system development process can have a positive influence on system implementation and better meet business expectations.

Application of Metrics in Early Stages of Process Development – Two Case Studies

AbstractZur Entwicklung nachhaltiger Prozesse ist es schon in frühen Phasen der Entwicklung sinnvoll, Methoden aus dem Bereich des betrieblichen Umwelt‐, Gesundheits‐ und Sicherheits‐Management (EHS‐Management) als auch aus dem Ökobilanzsektor zu nutzen. Vor dem Hintergrund einer geringen Datenverfügbarkeit und Datenqualität werden Möglichkeiten der Anwendung von Kennzahlen in den frühen Phasen der Prozessentwicklung an zwei Fallbeispielen vorgestellt. Zum einen wird eine vergleichende Stoffstromanalyse von zwei alternativen Syntheserouten zur Herstellung einer enantiomerenreinen β‐Aminosäure zur Bewertung von Gesundheits‐ und Kostenaspekten diskutiert. Zum anderen wird am Beispiel der Glutaraldehyd‐Produktion die Berechnung des kumulierten Energieaufwands mithilfe von Software‐Tools gezeigt.

SIMULATION OF PRODUCTS CLASSIFICATION SYSTEM FOR MANUFACTURING COST FORECASTING

This paper deals with product classification system, which is an essential factor to find proper product's performance and manufacturing cost, and potential manufacturer in production enterprises. The developed system can help classifying different products with similar purpose and functions on the separate class level in accordance of their characteristics and properties. The classification procedure is based on the data input at the interactive regime and applies attributes such as object mass, size, volume, power, speed and so on. For products that fall on the same class, the forecasting model of manufacturing cost has been developed. Forecasting model is grounded on the mathematical regression equation. The developed classification system and manufacturing resources forecasting model have been tested and validated in both laboratory and industry. The proposed research creations have been implemented in virtual manufacturing system and employed at the early stage of new product and process development stage. http://dx.doi.org/10.5755/j01.mech.17.3.507

Rapid High-Resolution Characterization of Functionally Important Monoclonal Antibody N-Glycans by Capillary Electrophoresis

Characterization of the N-glycosylation present in the Fc region of therapeutic monoclonal antibodies requires rapid, high-resolution separation methods to guarantee product safety and efficacy during all stages of process development. Determination of fucosylated oligosaccharides is particularly important during clone selection, product characterization, and lot release as fucose has been shown to adversely affect the ability of mAbs to induce antibody dependent cellular cytotoxicity (ADCC). Here, we apply a general capillary electrophoresis optimization strategy to separate functionally relevant fucosylated and afucosylated glycans on mononclonal antibody products in the presence of several high mannose oligosaccharides. The N-glycans chosen represent those most commonly reported on CHO cell derived therapeutic antibodies. A rapid (<7 min) high-resolution separation of 12 commonly reported and functionally important IgG glycans was developed by systematically evaluating the effects of selectivity (boric acid) and efficiency (linear polyacrylamide) enhancing additives. The approach can be used to rapidly optimize capillary electrophoresis separation of other glycan mixtures. Following optimization, the method was applied to overnight sample processing for automated 96 well plate-based glycosylation analyses of two nonproprietary therapeutic monoclonal antibodies, demonstrating ruggedness and suitability for high-throughput process and product monitoring applications.

Biotransformations in microstructured reactors: more than flowing with the stream?

The state of the art in the application of microstructured flow reactors for biocatalytic process research is reviewed. A microstructured reactor that is fully automated and analytically equipped presents a powerful screening tool with which to perform biocatalyst selection and optimization of process conditions at intermediary stages of process development. Enhanced mass transfer provided by the microstructured reactor can be exploited for process intensification, particularly during multiphase biocatalytic processing where mass transfer across phase boundaries is often limiting. Reversible immobilization of enzymes in microchannels remains a challenge for flexible realization of biotransformations in microstructured reactors. Compartmentalization in microstructured reactors could be useful in performing multistep chemoenzymatic conversions.

Detection of the II–I Etiracetam solvent-mediated polymorphic transformation through the online monitoring of the suspension apparent viscosity

Although going to complex techniques can be useful at some stage of process development of crystallization processes, the goal of this article is to show that process control can also be achieved with relatively simple measurements, depending of course on what type of control is needed. Focusing on the Etiracetam crystallization process, this work demonstrates the potential of the online monitoring of the power dissipated in the suspension, a property that can be directly related to the suspension apparent viscosity for the detection of the solvent-mediated polymorphic transformation from form II to form I. Even if the work presented in this paper focused on a specific crystallization process, this easy-to-use method seems promising for the control of other crystallization processes exhibiting solvent-mediated polymorphic transformations with similar characteristics: (i) significant difference in the PSD of the crystals of the stable and the metastable forms, (ii) solid volumetric fraction in the suspension higher than 40% and (iii) absence of phenomena (breaking, attrition, agglomeration, secondary nucleation, etc.) interfering with the polymorphic transformation and affecting the PSD of the crystals.

Analytical hierarchy processes (AHP) for the selection of solvents in early stages of pharmaceutical process development

The AHP (analytical hierarchy process) mathematical model was implemented into a tool aimed to aid the selection of solvents in the early stages of pharmaceutical process development. The tool assesses environmental implications using the information available in the early stages of development. Solvent properties, characteristics, and their relationship with common operations are exploited. In order to make the approach user-friendly, the tool was incorporated into a VB.NET application. The user obtains a ranked list of potentially good solvents. The result can be used as a starting point in solvent selection. The chemist can explore implications of the solvent selected not only from synthesis perspective, but also from an HSE perspective. A case study is presented for the replacement of benzene, where through a series of steps the chemist inputs ideal solvent characteristics and the importance of each characteristic in the decision. In this case solvent replacement is based in finding a solvent with the same solubility behaviour, but with less toxicity problems and at low cost. The tool considers a wide pool of solvents in a short time and produces ranked choices according to the chemist needs; taking account of both synthesis and HSE perspectives.

Secure E-commerce Web Development Framework

This study presents a framework for safety critical e-commerce application development based on an Extreme Programming methodology with inbuilt security across the development lifecycle to mitigate security lapses. This approach tightens security checks and balances at every stage of development process, minimizes any vulnerability that will manifest in production environment and avoids an unnecessary extension of the life cycle.

A novel conico-cylindrical flask aids easy identification of critical process parameters for cultivation of marine bacteria

A polymethylmethacrylate (PMMA) conico-cylindrical flask (CCF) with an inner arrangement consisting of eight equidistantly spaced rectangular strips mounted radially on a circular disk to provide additional surface area for microbial attachment and ports to allow air supply was employed for melanin production by Shewanella colwelliana and antibiotic production by Pseudoalteromonas rubra. The design allowed comparison of production between (1) CCF with hydrophobic surface (PMMA-CCF), (2) CCF with hydrophilic glass surface (GS-CCF), and (3) standard unbaffled Erlenmeyer flask (EF). Melanin production in the PMMA-CCF was higher by at most 33.5% and growth of S. colwelliana by at most 309.2% compared to the other vessels. Melanin synthesis was positively correlated with reactor surface area and hydrophobicity, suspended cell growth, and biofilm formation. Antibiotic production in the EF was higher by at most 83.3%, but growth of P. rubra was higher in the PMMA-CCF by at most 54.5% compared to the other vessels. A hydrophilic vessel surface, abundant air supply, but low shear stress enhanced antibiotic production. The CCF together with the EF allowed identification of the crucial parameters (vessel surface characteristics, growth, biofilm formation, and aeration) influencing productivity, knowledge of which in the initial stages of process development will facilitate informed decisions at the later phases.

Applications of 3D X-Ray Microscopy for Advanced Package Development

This paper describes the utilization of non-destructive imaging using 3D x-ray microscopy for package development and failure analysis. Four case studies are discussed to explain our methodology and its impact on our advanced packaging development effort. Identifying and locating failures embedded deep inside the package, such as a solder fatigue failure within a flip chip package, without the need for physical cross-sectioning is of substantial benefit because it preserves the package for further analysis. Also of utility is the ability to reveal the structural details of the package while producing superior quality 2D and volumetric images. The technique could be used not only for analysis of defects and failures, but also to characterize geometries and morphologies during the process and package development stage.

Scale‐down prediction of industrial scale pleated membrane cartridge performance

Flat-sheet membrane discs represent the current standard format used for experimental prediction of the scale-up of normal flow filtration processes. Use of this format is problematic, however, since the scale-down results typically show a 40-55% difference in performance compared to large-scale cartridges depending upon the feedstock used. In this work, novel pleated scale-down devices (Am=1.51-15.1×10(-3) m2) have been designed and fabricated. It is shown that these can more accurately predict the performance of industrial scale single-use pleated membrane cartridges (Am=1.06 m2) commonly used within biopharmaceutical manufacture. The single-use scale-down cartridges retain the same pleat characteristics of the larger cartridges, but require a reduced feed volume by virtue of a substantially diminished number of active membrane pleats. In this study, a 1,000-fold reduction in feed volume requirement for the scale-down cartridge with the smallest membrane area was achieved. The scale-down cartridges were tested both with clean water and a pepsin protein solution, showing flux-time relationships within 10% of the large-scale cartridge in both cases. Protein transmission levels were also in close agreement between the different scale cartridges. The similarity in performance of the scale-down and the large-scale cartridges, coupled with the low feed requirement, make such devices an excellent method by which rapid scale-up can be achieved during early stage process development for biopharmaceutical products. This new approach is a significant improvement over using flat-sheet discs as the quantitative similarity in performance with the large-scale leads to reliable scale-up predictions while requiring especially small volumes of feed material.

Packaging Challenges on Shrink Chip Technology

As technology grows, semiconductor industry evolved towards tinier & thinner packages to meet market demand. To create competitiveness, chip size shrinkage at front-end wafer fabrication is essential for newer packages technology. With the tireless effort from front-end fab, die size & thickness are reduced up to 30%. There are changes on wire bond pad as well, pad opening is reduced & pad metallization is changed to NiPPdAu with barrier metal underneath. This paper will discuss in details mainly on die bonding & wire bonding challenges for shrink chip technology. Critical defects such as vertical crack die, die surface contamination & passivation crack were observed during process development stage. Several efforts were made to address them accordingly. Finally this paper will share on new failure mechanism observed during ramp up, evaluations for risk assessment & future improvement ideas to overcome mass production obstacles.

Wafer Sort Bitmap Data Analysis Using the PCA-Based Approach for Yield Analysis and Optimization

Yield analysis is one of the most important subjects in IC companies. During the initial stage of new process development, several factors can greatly impact the yield simultaneously. Traditionally, several learning cycle iterations are required to solve yield loss issues. This paper describes a novel way to diagnose yield loss issues in less iteration. First, the failure classification of bitmap data is transferred to a new basis using principal component analysis. Second, the defective rates are calculated and the original bitmap data is reconstructed in the principal basis, allowing the yield loss space to be generated by Cluster Analysis. Third, physical failure analysis samples can be selected to solve yield loss issues. Furthermore, the new yield loss basis can be used to monitor the progress of yield improvement as a discriminate analysis measure for reducing failure patterns (bitmap failures).

Continuous screening system for inhibited enzyme catalysis: A membrane reactor approach

The screening of catalysts, substrates or conditions in the early stages of bioprocess development requires an enormous number of experiments and is a tedious, expensive and time-consuming task. Currently available screening systems can only be operated in batch or fed-batch mode, which can lead to severe misinterpretations of screening results. For example, catalysts that are inhibited by substrates or accumulating products will be excluded from further investigations in the early stages of process development despite the fact that they might be superior to other candidates in a different operational mode. Important and advantageous properties such as turnover stability can also be overshadowed by product inhibition. The aim of this study was to develop a novel screening system that enables continuous feeding of substrates and continuous removal of products. A prototype based on the membrane reactor concept was designed and operated for a model reaction, the hydrolysis of cellulose.

Dynamic Bayesian Evaluation Method for System Reliability Growth Based on In-Time Correction

Small sample size is often a limitation in reliability field tests for expensive, destructive systems, which makes it difficult to evaluate system reliability accurately. A new system's development process is usually comprised of several stages within which the system experiences design updating, and prototype field testing, thus enabling the improvement of system reliability. A dynamic Bayesian evaluation method is presented in this paper to cope with this problem for binomial systems whose reliability grows along with stages of the system's development process. To implement the proposed method, a new discrete reliability growth model, derived from the learning-curve property, is introduced to describe system reliability growth through the stages of the development process, which is also capable of predicting the reliability at the next stage, based on the data acquired in the current stage. The prediction result, which acts as the prior knowledge for reliability at the next stage, is applied to determine the prior distribution of system reliability through the Maximum Entropy method. The advantage of the method is that it employs field test data from various stages of the system's development process to evaluate system reliability, comparing with traditional methods. The given example demonstrates that the proposed method adapts to evaluate system reliability when the sample size of the field test is small.

Modeling electrostatic interactions of baculovirus vectors for ion-exchange process development

Product-related impurities constitute a major burden in the production of recombinant viral vectors for gene therapy and vaccination; it impairs not only the biological efficacy of the preparation but the process yield/productivity. Recombinant baculovirus was used as an enveloped virus model to address this issue. Given that ion-exchange chromatography is a process of choice for purification of viral vectors, the analysis of the electrostatic behavior can be instrumental for the improvement of impurity removal. The main species, product (infective virus particle) and product-derived impurities (dsDNA-, glycoprotein-, and envelope-deprived baculovirus particles), were isolated and correspondent ζ potentials were analyzed through dynamic light scattering. A model of the virus based on the viral components critical for biological function is proposed. The contribution of these viral components to the overall particle electrostatic interaction energy profile (calculated between the particle and a putative ion-exchange surface) was assessed as a function of ionic strength and pH. This resulted in a deterministic tool capable of distinguishing the electrostatic properties of the infective virus particle from the major virus-related impurities. Within an ion-exchange bind-elute process, this knowledge helps narrow the optimization space in early stage process development for viral vectors by predicting the best selectivity conditions.