Search results for: Stephane Lavirotte

Authors: Nathalie Stephenne, Mathieu Veschkens, Stéphane Palm, Christophe Charlemagne, Jacques Defoux

Abstract:

According to Europe’s first River basin Management Plans (RBMPs), 56% of European rivers failed to achieve the good status targets of the Water Framework Directive WFD. In Central European countries such as Belgium, even more than 80% of rivers failed to achieve the WFD quality targets. Although the RBMP’s should reduce the stressors and improve water body status, their potential to address multiple stress situations is limited due to insufficient knowledge on combined effects, multi-stress, prioritization of measures, impact on ecology and implementation effects. This paper describes a webGis prototype developed for the Walloon administration to improve the communication and the management of sediment dredging actions carried out in rivers and lakes in the frame of RBMPs. A large number of stakeholders are involved in the management of rivers and lakes in Wallonia. They are in charge of technical aspects (client and dredging operators, organizations involved in the treatment of waste…), management (managers involved in WFD implementation at communal, provincial or regional level) or policy making (people responsible for policy compliance or legislation revision). These different kinds of stakeholders need different information and data to cover their duties but have to interact closely at different levels. Moreover, information has to be shared between them to improve the management quality of dredging operations within the ecological system. In the Walloon legislation, leveling dredged sediments on banks requires an official authorization from the administration. This request refers to spatial information such as the official land use map, the cadastral map, the distance to potential pollution sources. The production of a collective geodatabase can facilitate the management of these authorizations from both sides. The proposed internet system integrates documents, data input, integration of data from disparate sources, map representation, database queries, analysis of monitoring data, presentation of results and cartographic visualization. A prototype of web application using the API geoviewer chosen by the Geomatic department of the SPW has been developed and discussed with some potential users to facilitate the communication, the management and the quality of the data. The structure of the paper states the why, what, who and how of this communication tool.

Keywords: sediments, web application, GIS, rivers management

Procedia PDF Downloads 405

Authors: Julia Cristina Bonaldo, Christophe L. Martin, Severine Romero Baivier, Stephane Mazerat

Abstract:

Alumina refractories are commonly used in steel and foundry industries. These refractories are prepared through a powder metallurgy route. They are a mixture of hard alumina particles and graphite platelets embedded into a soft carbonic matrix (binder). The powder can be cold pressed isostatically or uniaxially, depending on the application. The compact is then fired to obtain the final product. The quality of the product is governed by the microstructure of the composite and by the process parameters. The compaction behavior and the mechanical properties of the fired product depend greatly on the amount of each phase, on their morphology and on the initial microstructure. In order to better understand the link between these parameters and the macroscopic behavior, we use the Discrete Element Method (DEM) to simulate the compaction process and the fracture behavior of the fired composite. These simulations are coupled with well-designed experiments. Four mixes with various amounts of Al₂O₃ and binder were tested both experimentally and numerically. In DEM, each particle is modelled and the interactions between particles are taken into account through appropriate contact or bonding laws. Here, we model a bimodal mixture of large Al₂O₃ and small Al₂O₃ covered with a soft binder. This composite is itself mixed with graphite platelets. X-ray tomography images are used to analyze the morphologies of the different components. Large Al₂O₃ particles and graphite platelets are modelled in DEM as sets of particles bonded together. The binder is modelled as a soft shell that covers both large and small Al₂O₃ particles. When two particles with binder indent each other, they first interact through this soft shell. Once a critical indentation is reached (towards the end of compaction), hard Al₂O₃ - Al₂O₃ contacts appear. In accordance with experimental data, DEM simulations show that the amount of Al₂O₃ and the amount of binder play a major role for the compaction behavior. The graphite platelets bend and break during the compaction, also contributing to the macroscopic stress. Firing step is modeled in DEM by ascribing bonds to particles which contact each other after compaction. The fracture behavior of the compacted mixture is also simulated and compared with experimental data. Both diametrical tests (Brazilian tests) and triaxial tests are carried out. Again, the link between the amount of Al₂O₃ particles and the fracture behavior is investigated. The methodology described here can be generalized to other particulate materials that are used in the ceramic industry.

Keywords: cold compaction, composites, discrete element method, refractory materials, x-ray tomography

Procedia PDF Downloads 138

Authors: Alireza Fallahfard, Ludwig Vinches, Stephane Halle

Abstract:

In the workplace, the exposure level of airborne contaminants should be evaluated due to health and safety issues. It can be done by numerical models or experimental measurements, but the numerical approach can be useful when it is challenging to perform experiments. One of the simplest models is the well-mixed room (WMR) model, which has shown its usefulness to predict inhalation exposure in many situations. However, since the WMR is limited to gases and vapors, it cannot be used to predict exposure to aerosols. The main objective is to modify the WMR model to expand its application to exposure scenarios involving aerosols. To reach this objective, the standard WMR model has been modified to consider the deposition of particles by gravitational settling and Brownian and turbulent deposition. Three deposition models were implemented in the model. The time-dependent concentrations of airborne particles predicted by the model were compared to experimental results conducted in a 0.512 m3 chamber. Polystyrene particles of 1, 2, and 3 µm in aerodynamic diameter were generated with a nebulizer under two air changes per hour (ACH). The well-mixed condition and chamber ACH were determined by the tracer gas decay method. The mean friction velocity on the chamber surfaces as one of the input variables for the deposition models was determined by computational fluid dynamics (CFD) simulation. For the experimental procedure, the particles were generated until reaching the steady-state condition (emission period). Then generation stopped, and concentration measurements continued until reaching the background concentration (decay period). The results of the tracer gas decay tests revealed that the ACHs of the chamber were: 1.4 and 3.0, and the well-mixed condition was achieved. The CFD results showed the average mean friction velocity and their standard deviations for the lowest and highest ACH were (8.87 ± 0.36) ×10-2 m/s and (8.88 ± 0.38) ×10-2 m/s, respectively. The numerical results indicated the difference between the predicted deposition rates by the three deposition models was less than 2%. The experimental and numerical aerosol concentrations were compared in the emission period and decay period. In both periods, the prediction accuracy of the modified model improved in comparison with the classic WMR model. However, there is still a difference between the actual value and the predicted value. In the emission period, the modified WMR results closely follow the experimental data. However, the model significantly overestimates the experimental results during the decay period. This finding is mainly due to an underestimation of the deposition rate in the model and uncertainty related to measurement devices and particle size distribution. Comparing the experimental and numerical deposition rates revealed that the actual particle deposition rate is significant, but the deposition mechanisms considered in the model were ten times lower than the experimental value. Thus, particle deposition was significant and will affect the airborne concentration in occupational settings, and it should be considered in the airborne exposure prediction model. The role of other removal mechanisms should be investigated.

Keywords: aerosol, CFD, exposure assessment, occupational settings, well-mixed room model, zonal model

Procedia PDF Downloads 102

Authors: Dalicia Bouallouche, Jean-Baptiste Vioix, Stéphane Millot, Eric Busvelle

Abstract:

When researchers or logistics software developers deal with vehicle routing optimization, they mainly focus on minimizing the total travelled distance or the total time spent in the tours by the trucks, and maximizing the number of visited customers. They assume that the upstream real data given to carry the optimization of a transporter tours is free from errors, like customers’ real constraints, customers’ addresses and their GPS-coordinates. However, in real transporter situations, upstream data is often of bad quality because of address geocoding errors and the irrelevance of received addresses from the EDI (Electronic Data Interchange). In fact, geocoders are not exempt from errors and could give impertinent GPS-coordinates. Also, even with a good geocoding, an inaccurate address can lead to a bad geocoding. For instance, when the geocoder has trouble with geocoding an address, it returns those of the center of the city. As well, an obvious geocoding issue is that the mappings used by the geocoders are not regularly updated. Thus, new buildings could not exist on maps until the next update. Even so, trying to optimize tours with impertinent customers GPS-coordinates, which are the most important and basic input data to take into account for solving a vehicle routing problem, is not really useful and will lead to a bad and incoherent solution tours because the locations of the customers used for the optimization are very different from their real positions. Our work is supported by a logistics software editor Tedies and a transport company Upsilon. We work with Upsilon's truck routes data to carry our experiments. In fact, these trucks are equipped with TOMTOM GPSs that continuously save their tours data (positions, speeds, tachograph-information, etc.). We, then, retrieve these data to extract the real truck routes to work with. The aim of this work is to use the experience of the driver and the feedback of the real truck tours to validate GPS-coordinates of well geocoded addresses, and bring a correction to the badly geocoded addresses. Thereby, when a vehicle makes its tour, for each visited customer, the vehicle might have trouble with finding this customer’s address at most once. In other words, the vehicle would be wrong at most once for each customer’s address. Our method significantly improves the quality of the geocoding. Hence, we achieve to automatically correct an average of 70% of GPS-coordinates of a tour addresses. The rest of the GPS-coordinates are corrected in a manual way by giving the user indications to help him to correct them. This study shows the importance of taking into account the feedback of the trucks to gradually correct address geocoding errors. Indeed, the accuracy of customer’s address and its GPS-coordinates play a major role in tours optimization. Unfortunately, address writing errors are very frequent. This feedback is naturally and usually taken into account by transporters (by asking drivers, calling customers…), to learn about their tours and bring corrections to the upcoming tours. Hence, we develop a method to do a big part of that automatically.

Keywords: driver experience feedback, geocoding correction, real truck tours

Procedia PDF Downloads 674

Authors: Alexia Wu, Joel Ribis, Jean-Christophe Brachet, Emmanuel Clouet, Benoit Arnal, Elodie Rouesne, Stéphane Urvoy, Justine Roubaud, Yves Serruys, Frederic Lepretre

Abstract:

To enhance the safety of Light Water Reactor, accident tolerant fuel (ATF) claddings materials are under development. In the framework of CEA-AREVA-EDF collaborative program on ATF cladding materials, CEA has engaged specific studies on chromium coated zirconium alloys. Especially for Loss-of-Coolant-Accident situations, chromium coated claddings have shown some additional 'coping' time before achieving full embrittlement of the oxidized cladding, when compared to uncoated references – both tested in steam environment up to 1300°C. Nevertheless, the behavior of chromium coatings and the stability of the Zr-Cr interface under neutron irradiation remain unknown. Two main points are addressed: 1. Bulk Cr behavior under irradiation: Due to its BCC crystallographic structure, Cr is prone to Ductile-to-Brittle-Transition at quite high temperature. Irradiation could be responsible for a significant additional DBTT shift towards higher temperatures. 2. Zircaloy/Cr interface behavior under irradiation: Preliminary TEM examinations of un-irradiated samples revealed a singular Zircaloy-4/Cr interface with nanometric intermetallic phase layers. Such particular interfaces highlight questions of how they would behave under irradiation - intermetallic zirconium phases are known to be more or less stable under irradiations. Another concern is a potential enhancement of chromium diffusion into the zirconium-alpha based substrate. The purpose of this study is then to determine the behavior of such coatings after ion irradiations, as a surrogate to neutron irradiation. Ion irradiations were performed at the Jannus-Saclay facility (France). 20 MeV Kr8+ ions at 400°C with a flux of 2.8x1011 ions.cm-2.s-1 were used to irradiate chromium coatings of 1-2 µm thick on Zircaloy-4 sheets substrate. At the interface, the calculated damage is close to 10 dpa (SRIM, Quick Calculation Damage mode). Thin foil samples were prepared with FIB for both as-received and irradiated coated samples. Transmission Electron Microscopy (TEM) and in-situ tensile tests in a Scanning Electron Microscope are being used to characterize the un-irradiated and irradiated materials. High Resolution TEM highlights a great complexity of the interface before irradiation since it is formed of an alternation of intermetallic phases – C14 and C15. The interfaces formed by these intermetallic phases with chromium and zirconium show semi-coherency. Chemical analysis performed before irradiation shows some iron enrichment at the interface. The chromium coating bulk microstructures and properties are also studied before and after irradiation. On-going in-situ tensile tests focus on the capacity of chromium coatings to sustain some plastic deformation when tested up to 350°C. The stability of the Cr/Zr interface is shown after ion irradiation up to 10 dpa. This observation constitutes the first result after irradiation on these new coated claddings materials.

Keywords: accident tolerant fuel, HRTEM, interface, ion-irradiation

Procedia PDF Downloads 363

Authors: Stephane Maingard, Babu Paturi, Maura Daly, Finnola Armstrong

Abstract:

Background: Perinatal optimization involves the implementation of twelve intervention bundles of care at Our Lady of Lourdes Hospital, reliably delivering evidence-based interventions in the antenatal, intrapartum, and neonatal period to improve preterm outcomes. These key interventions (e.g. Antenatal steroids, Antenatal counselling, Optimal cord management, Respiratory management etc.) are based on WHO (World Health Organization, BAPM (British Association of Perinatal Medicine), and the latest 2022 European Consensus guidelines recommendations. Methodology: In February 2023, a quality improvement project team (pediatricians, neonatologists, obstetricians, clinical skills managers) was established, and a project implementation plan was developed. The Program Study Act implemented the following: 1. Antenatal consultation pathway, 2. Creation and implementation of a perinatal checklist for preterm births less than 34 weeks of gestation, 3. Process changes to ensure the checklist is completed, 4. Completion of parent and staff surveys, 5. Ongoing training. We collected and compared a range of data before and after implementation. Results: Preliminary analysis so far at 1 month demonstrates improvement in the following areas: 50% increase in antenatal counselling. Right place of birth increased from 85% to 100%. Magnesium sulphate increased from 56% to 100%. No change was observed in buccal colostrum administration (28%), delayed cord clamping (75%), caffeine administration (100%), blood glucose level at one hour of life > 2,6mmol (85%). There was also no change noted in respiratory support at resuscitation, CPAP only (47%), IPPV with CPAP (45%), IPPV with intubation (20%), and surfactant administration (28%). A slight decrease in figures was noted in the following: steroid administration from 80% to 75% and thermal care obtaining optimal temperature on admission (65% to 50%). Discussion: Even though the findings are preliminary, the directional improvement shows promise. Improved communication has been achieved between all stakeholders, including our patients, who are key team members. Adherence to the bundles of care will help to improve survival and neurodevelopmental outcomes as well as reduce the length of stay, thereby overall reducing the financial cost, considering the lifetime cost of cerebral palsy is estimated at €800,000 and reducing the length of stay can result in savings of up to €206,000. Conclusion: Preliminary results demonstrate improvements across a range of patient, process, staff, and financial outcomes. Our future goal is a seamless pathway of patient centered care for babies and their families. This project is an interdisciplinary collaboration to implement best practices for a vulnerable patient cohort. Our two main challenges are changing our organization’s culture as well as ensuring the sustainability of the project.

Keywords: perinatal, optimization, antenatal, counselling, IPPV

Procedia PDF Downloads 17

Authors: Matar Celine, Gaucel Sebastien, Gontard Nathalie, Guilbert Stephane, Guillard Valerie

Abstract:

To increase the very short shelf life of fresh fruits and vegetable, Modified Atmosphere Packaging (MAP) allows an optimal atmosphere composition to be maintained around the product and thus prevent its decay. This technology relies on the modification of internal packaging atmosphere due to equilibrium between production/consumption of gases by the respiring product and gas permeation through the packaging material. While, to the best of our knowledge, benefit of MAP for fresh fruits and vegetable has been widely demonstrated in the literature, its effect on shelf life increase has never been quantified and formalized in a clear and simple manner leading difficult to anticipate its economic and environmental benefit, notably through the decrease of food losses. Mathematical modelling of mass transfers in the food/packaging system is the basis for a better design and dimensioning of the food packaging system. But up to now, existing models did not permit to estimate food quality nor shelf life gain reached by using MAP. However, shelf life prediction is an indispensable prerequisite for quantifying the effect of MAP on food losses reduction. The objective of this work is to propose an innovative approach to predict shelf life of MAP food product and then to link it to a reduction of food losses and wastes. In this purpose, a ‘Virtual MAP modeling tool’ was developed by coupling a new predictive deterioration model (based on visual surface prediction of deterioration encompassing colour, texture and spoilage development) with models of the literature for respiration and permeation. A major input of this modelling tool is the maximal percentage of deterioration (MAD) which was assessed from dedicated consumers’ studies. Strawberries of the variety Charlotte were selected as the model food for its high perishability, high respiration rate; 50-100 ml CO₂/h/kg produced at 20°C, allowing it to be a good representative of challenging post-harvest storage. A value of 13% was determined as a limit of acceptability for the consumers, permitting to define products’ shelf life. The ‘Virtual MAP modeling tool’ was validated in isothermal conditions (5, 10 and 20°C) and in dynamic temperature conditions mimicking commercial post-harvest storage of strawberries. RMSE values were systematically lower than 3% for respectively, O₂, CO₂ and deterioration profiles as a function of time confirming the goodness of model fitting. For the investigated temperature profile, a shelf life gain of 0.33 days was obtained in MAP compared to the conventional storage situation (no MAP condition). Shelf life gain of more than 1 day could be obtained for optimized post-harvest conditions as numerically investigated. Such shelf life gain permitted to anticipate a significant reduction of food losses at the distribution and consumer steps. This food losses' reduction as a function of shelf life gain has been quantified using a dedicated mathematical equation that has been developed for this purpose.

Keywords: food losses and wastes, modified atmosphere packaging, mathematical modeling, shelf life prediction

Procedia PDF Downloads 182

Authors: Despoina Chochtoula, Aristidis Ilias, Yannis Stamatiou

Abstract:

Modbus is a protocol that enables the communication among devices which are connected to the same network. This protocol is, often, deployed in connecting sensor and monitoring units to central supervisory servers in Supervisory Control and Data Acquisition, or SCADA, systems. These systems monitor critical infrastructures, such as factories, power generation stations, nuclear power reactors etc. in order to detect malfunctions and ignite alerts and corrective actions. However, due to their criticality, SCADA systems are vulnerable to attacks that range from simple eavesdropping on operation parameters, exchanged messages, and valuable infrastructure information to malicious modification of vital infrastructure data towards infliction of damage. Thus, the SCADA research community has been active over strengthening SCADA systems with suitable data protection mechanisms based, to a large extend, on cryptographic methods for data encryption, device authentication, and message integrity protection. However, due to the limited computation power of many SCADA sensor and embedded devices, the usual public key cryptographic methods are not appropriate due to their high computational requirements. As an alternative, Elliptic Curve Cryptography has been proposed, which requires smaller key sizes and, thus, less demanding cryptographic operations. Until now, however, no such implementation has been proposed in the SCADA literature, to the best of our knowledge. In order to fill this gap, our methodology was focused on integrating Modbus, a frequently used SCADA communication protocol, with Elliptic Curve based cryptography and develop a server/client application to demonstrate the proof of concept. For the implementation we deployed two C language libraries, which were suitably modify in order to be successfully integrated: libmodbus (https://github.com/stephane/libmodbus) and ecc-lib https://www.ceid.upatras.gr/webpages/faculty/zaro/software/ecc-lib/). The first library provides a C implementation of the Modbus/TCP protocol while the second one offers the functionality to develop cryptographic protocols based on Elliptic Curve Cryptography. These two libraries were combined, after suitable modifications and enhancements, in order to give a modified version of the Modbus/TCP protocol focusing on the security of the data exchanged among the devices and the supervisory servers. The mechanisms we implemented include key generation, key exchange/sharing, message authentication, data integrity check, and encryption/decryption of data. The key generation and key exchange protocols were implemented with the use of Elliptic Curve Cryptography primitives. The keys established by each device are saved in their local memory and are retained during the whole communication session and are used in encrypting and decrypting exchanged messages as well as certifying entities and the integrity of the messages. Finally, the modified library was compiled for the Android environment in order to run the server application as an Android app. The client program runs on a regular computer. The communication between these two entities is an example of the successful establishment of an Elliptic Curve Cryptography based, secure Modbus wireless communication session between a portable device acting as a supervisor station and a monitoring computer. Our first performance measurements are, also, very promising and demonstrate the feasibility of embedding Elliptic Curve Cryptography into SCADA systems, filling in a gap in the relevant scientific literature.

Keywords: elliptic curve cryptography, ICT security, modbus protocol, SCADA, TCP/IP protocol

Procedia PDF Downloads 270

Authors: Azeem Tariq, Stephane De Tourdonnet, Lars Stoumann Jensen, Reiner Wassmann, Bjoern Ole Sander, Quynh Duong Vu, Trinh Van Mai, Andreas De Neergaard

Abstract:

Rice growing area is increasing to meet the food demand of increasing population. Mostly, rice is growing on lowland, small landholder fields in most part of the world, which is one of the major sources of greenhouse gases (GHG) emissions from agriculture fields. The strategies such as, altering water and residues (carbon) management practices are assumed to be essential to mitigate the GHG emissions from flooded rice system. The actual implementation and potential of these measures on small farmer fields is still challenging. A field study was conducted on red river delta in Northern Vietnam to identify the potential challenges and barriers to the small rice farmers for implementation of climate smart rice practices. The objective of this study was to develop and access the feasibility of climate smart rice prototypes under actual farmer conditions. Field and scientific oriented framework was used to meet our objective. The methodological framework composed of six steps: i) identification of stakeholders and possible options, ii) assessment of barrios, drawbacks/advantages of new technologies, iii) prototype design, iv) assessment of mitigation potential of each prototype, v) scenario building and vi) scenario assessment. A farm survey was conducted to identify the existing farm practices and major constraints of small rice farmers. We proposed the two water (pre transplant+midseason drainage and early+midseason drainage) and one straw (full residue incorporation) management option keeping in views the farmers constraints and barriers for implementation. To test new typologies with existing prototypes (midseason drainage, partial residue incorporation) at farmer local conditions, a participatory field experiment was conducted for two consecutive rice seasons at farmer fields. Following the results of each season a workshop was conducted with stakeholders (farmers, village leaders, cooperatives, irrigation staff, extensionists, agricultural officers) at local and district level to get feedbacks on new tested prototypes and to develop possible scenarios for climate smart rice production practices. The farm analysis survey showed that non-availability of cheap labor and lacks of alternatives for straw management influence the small farmers to burn the residues in the fields except to use for composting or other purposes. Our field results revealed that application of early season drainage significantly mitigates (40-60%) the methane emissions from residue incorporation. Early season drainage was more efficient and easy to control under cooperate manage system than individually managed water system, and it leads to both economic (9-11% high rice yield, low cost of production, reduced nutrient loses) and environmental (mitigate methane emissions) benefits. The participatory field study allows the assessment of adaptation potential and possible benefits of climate smart practices on small farmer fields. If farmers have no other residue management option, full residue incorporation with early plus midseason drainage is adaptable and beneficial (both environmentally and economically) management option for small rice farmers.

Keywords: adaptation, climate smart agriculture, constrainsts, smallholders

Procedia PDF Downloads 266

Authors: Rowan Moore, Kai Scheffler, Eugen Damoc, Jennifer Sutton, Aaron Bailey, Stephane Houel, Simon Cubbon, Jonathan Josephs

Abstract:

Purpose: MS analysis of monoclonal antibodies (mAbs) at the protein and peptide levels is critical during development and production of biopharmaceuticals. The compositions of current generation therapeutic proteins are often complex due to various modifications which may affect efficacy. Intact proteins analyzed by MS are detected in higher charge states that also provide more complexity in mass spectra. Protein analysis in native or native-like conditions with zero or minimal organic solvent and neutral or weakly acidic pH decreases charge state value resulting in mAb detection at higher m/z ranges with more spatial resolution. Methods: Three commercially available mAbs were used for all experiments. Intact proteins were desalted online using size exclusion chromatography (SEC) or reversed phase chromatography coupled on-line with a mass spectrometer. For streamlined use of the LC- MS platform we used a single SEC column and alternately selected specific mobile phases to perform separations in either denaturing or native-like conditions: buffer A (20 % ACN, 0.1 % FA) with Buffer B (100 mM ammonium acetate). For peptide analysis mAbs were proteolytically digested with and without prior reduction and alkylation. The mass spectrometer used for all experiments was a commercially available Thermo Scientific™ hybrid Quadrupole-Orbitrap™ mass spectrometer, equipped with the new BioPharma option which includes a new High Mass Range (HMR) mode that allows for improved high mass transmission and mass detection up to 8000 m/z. Results: We have analyzed the profiles of three mAbs under reducing and native conditions by direct infusion with offline desalting and with on-line desalting via size exclusion and reversed phase type columns. The presence of high salt under denaturing conditions was found to influence the observed charge state envelope and impact mass accuracy after spectral deconvolution. The significantly lower charge states observed under native conditions improves the spatial resolution of protein signals and has significant benefits for the analysis of antibody mixtures, e.g. lysine variants, degradants or sequence variants. This type of analysis requires the detection of masses beyond the standard mass range ranging up to 6000 m/z requiring the extended capabilities available in the new HMR mode. We have compared each antibody sample that was analyzed individually with mixtures in various relative concentrations. For this type of analysis, we observed that apparent native structures persist and ESI is benefited by the addition of low amounts of acetonitrile and formic acid in combination with the ammonium acetate-buffered mobile phase. For analyses on the peptide level we analyzed reduced/alkylated, and non-reduced proteolytic digests of the individual antibodies separated via reversed phase chromatography aiming to retrieve as much information as possible regarding sequence coverage, disulfide bridges, post-translational modifications such as various glycans, sequence variants, and their relative quantification. All data acquired were submitted to a single software package for analysis aiming to obtain a complete picture of the molecules analyzed. Here we demonstrate the capabilities of the mass spectrometer to fully characterize homogeneous and heterogeneous therapeutic proteins on one single platform. Conclusion: Full characterization of heterogeneous intact protein mixtures by improved mass separation on a quadrupole-Orbitrap™ mass spectrometer with extended capabilities has been demonstrated.

Keywords: disulfide bond analysis, intact analysis, native analysis, mass spectrometry, monoclonal antibodies, peptide mapping, post-translational modifications, sequence variants, size exclusion chromatography, therapeutic protein analysis, UHPLC

Procedia PDF Downloads 361