Towards continuous biomanufacturing a computational approach for the intensification of monoclonal antibody production

Current industrial trends encourage the development of sustainable, environmentally friendly processes with reduced energy and raw material consumption. Meanwhile, the increasing market demand as well as the tight regulations in product quality, necessitate efficient operating procedures that guarantee products of high purity. In this direction, process intensification via continuous operation paves the way for the development of novel, eco-friendly processes, characterized by higher productivity compared to batch (Nicoud, 2014). The shift towards continuous operation could advance the market of high value biologics, such as monoclonal antibodies (mAbs), as it would lead to shorter production times, decreased costs, as well as significantly less energy consumption (Konstantinov and Cooney, 2015, Xenopoulos, 2015). In particular, mAb production comprises two main steps: the culturing of the cells (upstream) and the purification of the targeted product (downstream). Both processes are highly complex and their performance depends on various parameters. In particular, the efficiency of the upstream depends highly on cell growth and the longevity of the culture, while product quality can be jeopardized in case the culture is not terminated timely. Similarly, downstream processing, whose main step is the chromatographic separation, relies highly on the setup configuration, as well as on the composition of the upstream mixture. Therefore, it is necessary to understand and optimize both processes prior to their integration. In this direction, the design of intelligent computational tools becomes eminent. Such tools can form a solid basis for the: (i) execution of cost-free comparisons of various operating strategies, (ii) design of optimal operation profiles and (iii) development of advanced, intelligent control systems that can maintain the process under optimal operation, rejecting disturbances. In this context, this work focuses on the development of advanced computational tools for the improvement of the performance of: (a) chromatographic separation processes and (b) cell culture systems, following the systematic PAROC framework and software platform (Pistikopoulos et al., 2015). In particular we develop model-based controllers for single- and multi-column chromatographic setups based on the operating principles of an industrially relevant separation process. The presented strategies are immunized against variations in the feed stream and can successfully compensate for time delays caused due to the column residence time. Issues regarding the points of integration in multi-column systems are also discussed. Moreover, we design and test in silico model-based control strategies for a cell culture system, aiming to increase the culture productivity and drive the system towards continuous operation. Challenges and potential solutions for the seamless integration of the examined bioprocess are also investigated at the end of this thesis.Open Acces

Parents-Teachers Transformational Community of Inquiry: An Innovative Model

ABSTRACT: Extensive research over the last 3 decades has documented that collaboration between parents and teachers in the education of children from kindergarten through high school is highly beneficial for all parties, and particularly so for the academic progress and general welfare of the students. This dissertation describes a study that was directed toward developing and refining an innovative model of teaching practice—Philosophy for parent (P4P)—specifically directed at the parents of K-12 school-age children for the purpose of bridging a now widely recognized gap in communication and collaboration between parents and teachers—and hence in potential educational efficacy—and convert the actors into a well-bonded community—one that is actively engaged via "philosophic" Socratic dialogue with questions that probe and challenge their own and mutual frames of reference. And one that will intentionally sponsor and facilitate parents' direct involvement with their children's curriculum and everyday progress. In particular, my research focused on creating an innovative model of practice directed toward promoting teacher-facilitated liberating democratic dialogue between parents and teachers that is based on the fundamental principles of Socratic dialogue that academically acclaimed Columbia University professor of philosophy, Matthew Lipman, incorporated into an educational model for schoolteachers, and their students. In Lipman's model the teacher fosters and facilitates formation of a Community of Philosophical Inquiry among students from preschool age to adolescence. Use of this model—termed Philosophy for/with Children, or P4C—has been found in research (Lipman et al., 1980, p.15, Vansielegheim & Kennedy, 2011) to upgrade and cultivate children's innate abilities in questioning, reflecting, and thinking critically in a liberating environment if they are given appropriate prompts at school and home. Use of the process of philosophical Socratic dialogue—not study of historical philosophy—to explore and evaluate alternative actions and concepts between both children and adults is highly innovative in light of the fact that, until very recently, it was unimagined that children could think abstractly let alone compare and balance philosophical concepts. I call my model in which teachers employ adult-adapted versions of Lipman's principles, Philosophy for Parents (P4P). In time I perceived under intensive study of the literature that a number of adult learning theories, especially including Mezirow's Transformative Learning and its dialogue circles, and, to some extent others, might likely prove, under study in practice, to elucidate and positively inform the process and evolution of P4P. My P4P model was developed and pilot tested in a 2-year qualitative Action Research Study in a Kindergarten in Greece—with observed favorable results. During study of underlying theory at University of Naples Federico II, I decided to test P4P in the United States where Lipman's P4C has been admired, practiced, and extensively studied. However, while the first meetings with parents in participating school sites and were in progress—the study was rapidly terminated by an unforeseen Pandemic caused by worldwide, rapidly proliferating Covid-19 disease and attendant discontinuance of face-to-face teaching in schools. And so, an alternative path of study had to be devised. In concert, my doctoral study sponsor at the University of Naples Federico II, Maura Striano, and I converted my Action-Research-in-practice study to a conceptual study of my P4P model refined by extensive further research into its theoretic substrate—augmented by a complementary 49-question, self-administered, comparative international survey of a snowball sample of parents of schoolchildren from generally high SES and academically inclined families in the United States, Italy, Greece, Spain and Costa Rica. The survey's purpose was to examine subject interest in and direct involvement with their child/ren's education, their evaluation of the impact of relationships within the family and with their child/ren's school on their academic and general future success in life, and their appreciation of factors that might signal receptiveness to engaging with my model if it were offered in their child/ren's school. In a chrono-situational context, it probed parents' level of, and interest in, engagement with their children's school both before and then during Covid-19 remote learning, and comparison of the strengths of any factors that enhanced their interest and engagement with the school under pre-Covid-19 vs. pandemic conditions. While it is quite unlikely that many, or even any of the parents in the sample knew anything even about P4C, responses did at least suggest their likely inclination toward P4P—among them, surprisingly, a preference for dialogic communication. An ancillary purpose pursued in this period was development of a Teacher's Handbook as a tool that would assist teachers in leading this practice, featuring stories that might under the right conditions be expected to provoke free and open dialogic discussion between almost any possible grouping of parents and teacher-facilitators, where judgment of "right" or "wrong" might often be suspended to the point of irrelevance, where there might be few, if any, a priori "given" answers ever sanctioned, and where it would be encouraged that one person's point would be commented on, such that one person's question is quite likely to be "answered" (i.e., responded to) by another person's question. Evolution of a teacher-facilitated Community of Philosophical Inquiry, aspirationally, might be expected to eventuate in meaningful parent-teacher partnerships where all actors think deeply, exchange ideas, argue with respect to their own and others' voices and build relevant capacities to manage the complexity of modern pedagogy together as allies on a team. Abstract di Tesi Un vasto filone di ricerca negli ultimi 3 decenni ha documentato che la collaborazione tra genitori e insegnanti nell'educazione dei bambini, dalla scuola materna alla scuola superiore, è altamente vantaggiosa per tutte le parti e, in particolare, per il progresso scolastico e il benessere generale degli studenti. Questa tesi descrive uno studio che è stato diretto allo sviluppo e al perfezionamento di un modello innovativo di pratica didattica - Filosofia per i genitori (Philosophy for Parents, P4P) - rivolto nello specifico ai genitori di bambini in età scolare K-12 (primaria e secondaria) allo scopo di colmare un divario ora ampiamente riconosciuto nella comunicazione e collaborazione tra genitori e insegnanti - e quindi nella potenziale efficacia educativa. Esso trasforma gli attori in una comunità ben legata, attivamente coinvolta attraverso il dialogo socratico "filosofico" con domande che sondano e sfidano i propri e reciproci quadri di riferimento . Inoltre, esso che sponsorizzerà intenzionalmente e faciliterà il coinvolgimento diretto dei genitori nel curriculum dei loro figli e il loro progresso quotidiano. In particolare, la mia ricerca si è concentrata sulla creazione di un modello di pratica innovativo, volto a promuovere il dialogo democratico liberatorio tra genitori e insegnanti, facilitato dall'insegnante, basato sui principi fondamentali del dialogo socratico che il celebre professore di filosofia dalla Columbia University, Matthew Lipman, ha incorporato in un modello educativo per gli insegnanti e i loro studenti. Nel modello di Lipman, l'insegnante promuove e facilita la formazione di una comunità di indagine filosofica tra gli studenti, dall'età prescolare all'adolescenza. La ricerca (Lipman et al., 1980, p.15, Vansielegheim & Kennedy, 2011) ha mostrato come l'uso di questo modello, chiamato Filosofia per/con i bambini, o P4C, migliori e coltivi le abilità innate dei bambini nel mettere in discussione, riflettere, e pensare in modo critico in un ambiente liberatorio, se gli vengono dati suggerimenti appropriati a scuola e a casa. L'uso del processo del dialogo filosofico socratico, e non lo studio della filosofia storica, per esplorare e valutare azioni e concetti alternativi tra bambini e adulti è altamente innovativo alla luce del fatto che, fino a tempi molto recenti, era impensabile che i bambini potessero pensare in modo astratto, figurarsi confrontare e bilanciare concetti filosofici. Chiamo il mio modello, in cui gli insegnanti utilizzano versioni adattate per adulti dei principi di Lipman, Filosofia per i genitori (P4P). Col tempo ho percepito, attraverso uno studio intensivo della letteratura, che un certo numero di teorie sull'apprendimento degli adulti, tra cui in particolare l'apprendimento trasformativo di Mezirow e i suoi circoli di dialogo, e in una certa misura altre, potrebbero probabilmente dimostrare, allo studio pratico, di chiarire e informare positivamente il processo ed evoluzione del P4P. Il mio modello P4P è stato sviluppato e testato in uno studio pilota qualitativo, di ricerca- azione, di 2 anni in un asilo in Grecia, con risultati favorevoli osservati. Durante lo studio della teoria all'Università degli Studi di Napoli Federico II, ho deciso di testare il P4P negli Stati Uniti, dove il P4C di Lipman è stato ammirato, praticato e ampiamente studiato. Tuttavia, mentre erano in corso i primi incontri con i genitori nelle sedi scolastiche aderenti, lo studio è stato rapidamente sospeso da una pandemia imprevista causata dalla malattia Covid-19, in rapida proliferazione in tutto il mondo, e dalla conseguente interruzione dell'insegnamento faccia a faccia nelle scuole. E così è stato necessario ideare un percorso di studio alternativo. Insieme, la mia tutor di dottorato presso l'Università degli Studi di Napoli Federico II, Maura Striano, ed io abbiamo convertito il mio studio empirico di ricerca-azione in uno studio concettuale del mio modello P4P, perfezionato da ulteriori approfondite ricerche sul suo substrato teorico e ampliato da un questionario comparativo internazionale di 49 domande, autosomministrato , ad un campione a valanga di genitori di bambini in età scolare di status socio-economico generalmente alto e di famiglie con tendenze accademiche negli Stati Uniti, in Italia, in Grecia, in Spagna e in Costa Rica. Lo scopo dell'indagine era di esaminare l'interesse dei soggetti e il coinvolgimento diretto nell'istruzione dei figli, la loro valutazione dell'impatto delle relazioni all'interno della famiglia e con la scuola sul loro successo scolastico e futuro generale nella vita, e la loro valutazione dei fattori che potrebbero segnalare la ricettività a impegnarsi con il mio modello se fosse offerto nella scuola del figlio . In un contesto crono-situazionale, il questionario ha sondato il livello di coinvolgimento e l'interesse dei genitori nei confronti della scuola dei propri figli, sia prima che durante l'apprendimento a distanza da Covid-19, e il confronto dei punti di forza di tutti i fattori che hanno accresciuto il loro interesse e il loro coinvolgimento nei confronti della scuola in condizioni pre-Covid-19 vs in pandemia. Anche se è abbastanza improbabile che molti, o anche qualcuno dei genitori nel campione, sapesse qualcosa del P4C, le risposte suggerivano la loro probabile inclinazione verso il P4P, tra cui, sorprendentemente, una preferenza per la comunicazione dialogica. Uno scopo accessorio perseguito in questo periodo è stato lo sviluppo di un Manuale dell'insegnante, come strumento che avrebbe aiutato gli insegnanti a condurre questa pratica, con storie che, nelle giuste condizioni, possano provocare discussioni dialogiche libere e aperte tra quasi tutti i possibili gruppi di genitori e docenti-facilitatori, dove il giudizio di "giusto" o "sbagliato" possa essere spesso sospeso fino all'irrilevanza, dove possano esserci poche, se non nessuna, risposte "date a priori" e mai approvate, e dove possa essere incoraggiato il commentare il punto di una persona , in modo tale da rendere molto probabile che la domanda di una persona "riceva una risposta" dalla domanda di un'altra persona. Ci si potrebbe aspettare che l'evoluzione di una comunità di indagine filosofica facilitata dall'insegnante sfoci in partnership genitori-insegnanti significativa, in cui tutti gli attori pensano profondamente, scambiano idee, discutono rispetto alla propria voce e a quella degli altri e costruiscono capacità rilevanti per gestire la complessità della pedagogia moderna insieme, come alleati, in squadra

CAR T-cell therapies: The concept of a dynamic supply chain

Novarti’s Kymriah and Kite’s Yescarta, are the first Chimeric Antigen Receptor (CAR) T-cell therapies to receive regulatory approval both in the United States and in Europe. They are suggested as new face in cancer treatment and in particular in B-cell acute lymphoblastic leukaemia (ALL). Their promising results have encouraged numerous research groups and manufacturers to explore the potential of those therapies in the treatment of various cancer types, resulting into 317 clinical trials globally (based on a recent search on ClinicalTrials.gov (2018)). Today, in the UK CAR T cells are only available through clinical trial schemes (approximately 250 patients per trial), thus being produced and delivered at a small scale. However, their recent European Medicines Agency Approval will allow them to become available to a wider patient population (approximately 40,000 eligible patients by 2031 (Figure 1), based on research performed on the UK patient population), requiring, therefore significant scale up/out both in the manufacturing line as well as in the logistics/supply chain model. In this work we focus on the design of a modelling tool to assist the decision making in the design of the supply chain model of CAR T cell therapies. Expanding our previous work (Wang et al., 2018), we demonstrate the design of a Resource Technology Network (RTN) for the identification of the key steps/decisions in the CAR T supply chain model. Based on previous qualitative results (Papathanasiou, 2018), we present a comparison between three supply chain model structures and we introduce the concept of the “dynamic” supply chain model. The latter refers to a versatile supply chain network that is tailored to the varying therapy demand. Based on the demand profiles, the modelling tool decides on: (a) number and location of clinical sites, (b) number and location of manufacturing sites and (c) best means of transport for the therapy. Lastly, the model considers time restrictions related to product shelf life, as well as different business decisions (e.g. in-house versus outsourcing quality control). Please click Additional Files below to see the full abstract

parents' philosophical community: when parents go to school!

Research seems to be explicit on children's benefit from parent's participation in their schooling. The ways, though, parents can be involved are not yet apparent. A variety of educational strategies and programs are being tested globally in order to enhance the collaboration of the school with the family. Through Action Research, the effectiveness of an initiative of cooperation with the parents in a kindergarten school in Athens has been explored, during the School Years 2014-15 and 2015-16. The successful engagement of philosophical practices for the students in previous years exposed the need for adopting the specific tool of Philosophy for Children for the community of parents. Carefully selected stories have been used as a stimulus for raising philosophical questions that were analysed in depth by parents and teachers. The creation of a philosophical community of inquiry with parents within school, revealed a new horizon of communication and cooperation while raising respect and empathy among all participants. This research exhibits that building communities of philosophical inquiry with children and adults likewise, and within the same environment reflects and strengthens dialogue between all parts of the school community triangle (students, teachers, parents) and it therefore seems to be an effective approach to be implemented

A model-based approach towards accelerated process development: A case study on chromatography

Process development is typically associated with lengthy wet-lab experiments for the identification of good candidate setups and operating conditions. In this paper, we present the key features of a model-based approach for the identification and assessment of process design space (DSp), integrating the analysis of process performance and flexibility. The presented approach comprises three main steps: (1) model development & problem formulation, (2) DSp identification, and (3) DSp analysis. We demonstrate how such an approach can be used for the identification of acceptable operating spaces that enable the assessment of different operating points and quantification of process flexibility. The above steps are demonstrated on Protein A chromatographic purification of antibody-based therapeutics used in biopharmaceutical manufacturing.Comment: Pre-print paper under revie

TELLING TRANSVERSAL COMPETENCES TO BE PROFESSIONALLY PROMOTED

This article introduces experimental reflection on the experiences at an ‘Employability Skills’ laboratory1 of a group of young volunteers from the National Civilian Service under the ‘Support and Inclusion’ project of the Employment Promotion Section (SPO in Italian) of the University of Naples Federico II SInAPSi Centre. Young volunteers were included as unstructured support figures in activities that sought mainly to assist and serve students with disabilities. More specifically, these activities included: accompaniment and support during lessons; digitization of teaching material; providing support for the assorted services offered by the University Centre; general training implemented by AMESCI staff; specific training implemented by SInAPSi operatives. The experiences, which were accomplished in cooperation with the Europe 2020 programme, included the creation of an integrated system to recognize and validate formal, non-formal, and informal skills, as a tool to promote youth employment (Striano, Capobianco 2016)

Chapter Guidance Models and Practices Adopted Internationally to Promote the Exploration of Skills Relating to the Employability of Students with Disabilities. A First Meta-Analysis

Employability is defined as an interweaving of a person’s human, social and psychological capital, mediated by situational variables, which allows individuals to enter the job market with a professional personal project (Grimaldi, Porcelli, Rossi 2014). Nowadays, young people enter the job market through long, precarious, and poorly contextualized paths, while the socialization processes become recursive, discontinuous, and fragmented (Lodigiani 2010). A key role can be played by guidance services, which can start at university, to meet the demands of the (many) young people who are discouraged and disillusioned to the point where they cannot even imagine a job while still at university. In the employability stakes, what is even more complex is the encounter between young people with disabilities and the world of work, due to the persistence of stereotypes and stigmas. Research questions: What are the intervention models and guidance practices adopted by university guidance services internationally to promote the exploration of skills relating to the employability of students with disabilities? Objectives: To analyse the main intervention models and guidance practices adopted internationally to explore the skills associated with employability in students with disabilities. Methodology: It was decided to carry out a theoretical analysis of 20 scientific articles concerning the models and practices adopted to explore the competences relating to employability in certain university orientation services for students with disabilities in Italy, France, the UK, and the United States. NVivo software was used (Richards 1999) to systematically explore the scientific literature. Preliminary Findings: A first scientific paper showed that, like in Italy and France, the «Competence Balance Sheet» (Ardouin 2010) is the guiding practice in the USA, while in the UK, it is the Career Guidance Approach (Reid, Scott 2010). In the literature, orientation models and practices are also closely linked to the various patterns of employability. Final remarks: The implementation of guidance counseling paths aimed at exploring the skills associated with employability among all students and graduates is crucial to the completion of a viable strategic action in the University’s social function, as a part of new organizational models that take the plurality of learning opportunities into accoun

Advanced computational tools to enhance continuous monoclonal antibody production

Leading pharmaceutical companies invest high percentage of their revenue in the improvement of existing technologies used for the production of monoclonal antibodies (mAbs). Recently, there has been a paradigm shift towards the development of continuous/quasi-continuous purification operations, aiming to reduce capital and operational costs [1]. At the moment, however, there are no standardized methods and/or tools that can be used for global control and monitoring of integrated processes. Mathematical models and advanced computational tools can be the key for the development of robust, integrated processes, as they can provide valuable insight in the process dynamics and ensure optimal operation [2]. However, such processes are usually characterized by complex mathematical models and periodic operation profiles that result into computationally expensive solutions and challenge the development of global control methods and tools. In this work, we are presenting a novel approach for the development of advanced controllers towards the intensification of mAb production, considering the fed-batch culturing of GS-NS0 cells and the semi-continuous Multicolumn Countercurrent Solvent Gradient Purification (MCSGP) process [3]. The controller development is realized via the application of a generic framework for the development of advanced control strategies (PAROC) [4] that involves: (i) development of a high-fidelity process model, (ii) approximation of the complex, process model, (iii) design of the multi-parametric controller, (iv) ‘closed-loop’, in-silico validation of the controller against the process model. The development of the control policies is based on multi-parametric Model Predictive Control (mp-MPC) policies that reduce the online, computational force of the controller by deriving the control inputs as a set of explicit functions of the system states and can be implemented on embedded devices [5]. One of the main advantages of the proposed framework is the ability to test the controllers ‘in-silico’, against the high-fidelity process model and evaluate their performance before operating them online. The results from this study indicate that optimal operation, under maximum purity and productivity yield can be ensured with the development of advanced computational tools. The control policies are applied both in the upstream and the downstream processing; yielding therefore a fertile ground towards the development of a global control strategy that can ensure continuous operation

Chapter Telling Transversal Competences… to be Professionally Promoted

This article introduces experimental reflection on the experiences at an ‘Employability Skills’ laboratory of a group of young volunteers from the National Civilian Service under the ‘Support and Inclusion’ project of the Employment Promotion Section (SPO in Italian) of the University of Naples Federico II SInAPSi Centre. Young volunteers were included as unstructured support figures in activities that sought mainly to assist and serve students with disabilities. More specifically, these activities included: accompaniment and support during lessons; digitization of teaching material; providing support for the assorted services offered by the University Centre; general training implemented by AMESCI staff; specific training implemented by SInAPSi operatives. The experiences, which were accomplished in cooperation with the Europe 2020 programme, included the creation of an integrated system to recognize and validate formal, non-formal, and informal skills, as a tool to promote youth employment (Striano, Capobianco 2016

Operability-economics trade-offs in adsorption-based CO2_2 capture process

Low-carbon dispatchable power underpins a sustainable energy system, providing load balancing complementing wide-scale deployment of intermittent renewable power. In this new context, fossil fuel-fired power plants must be coupled with a post-combustion carbon capture (PCC) process capable of highly transient operation. To tackle design and operational challenges simultaneously, we have developed a computational framework that integrates process design with techno-economic assessment. The backbone of this is a high-fidelity PCC mathematical model of a pressure-vacuum swing adsorption process. We demonstrate that the cost-optimal design has limited process flexibility, challenging reactiveness to disturbances, such as those in the flue gas feed conditions. The results illustrate that flexibility can be introduced by relaxing the CO$_2$ recovery constraint on the operation, albeit at the expense of the capture efficiency of the process. We discover that adsorption-based processes can accommodate for significant flexibility and improved performance with respect to the operational constraints on CO$_2$ recovery and purity. The results herein demonstrate a trade-off between process economics and process operability, which must be effectively rationalised to integrate CO$_2$ capture units in the design of low-carbon energy systems.Comment: Pre-print paper currently under review. 32 pages, 6 figures. The first two authors contributed equally to this wor