Advanced monitoring & control in microbial cultivation processes for recombinant protein production [Elektronische Ressource] / von Marco Jenzsch

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• Publié par: martin-luther-universitat_halle-wittenberg
• Publié le: 01 janvier 2006
• Nombre de lectures: 48
• Langue: Deutsch
• Poids de l'ouvrage: 3 Mo

Extrait

Advanced Monitoring & Control
in Microbial Cultivation Processes
for Recombinant Protein Production




Dissertation

zur Erlangung des akademischen Grades

Doktoringenieur (Dr.-Ing.)


genehmigt durch die


Mathematisch-Naturwissenschaftlich-Technische Fakultät
- Ingenieurwissenschaftlicher Bereich -
der Martin-Luther-Universität Halle-Wittenberg


von

Herrn Dipl.-Ing. Marco Jenzsch

geb. am 09. Dezember 1978 in Merseburg




Dekan der Fakultät: Prof. Dr. rer. nat. habil. H. Graener


Gutachter: Prof. Dr. rer. nat. habil. A. Lübbert
Prof. em. Dr. Dr. h.c. K. Schügerl


Halle (Saale), den 13. Juli 2006
urn:nbn:de:gbv:3-000010501
[http://nbn-resolving.de/urn/resolver.pl?urn=nbn%3Ade%3Agbv%3A3-000010501]
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Danksagung


Diese Arbeit wurde im Zeitraum von November 2002 bis Mai 2006 am Zentrum für
Bioverfahrenstechnik der Martin-Luther-Universität Halle-Wittenberg in der Arbeitsgruppe
von Herrn Prof. Dr. Andreas Lübbert angefertigt.
Ich danke allen Mitarbeitern für das angenehme Arbeitsklima und die Unterstützung bei den
verschiedensten Problemstellungen die sich während dieser Zeit ergaben.
Für eine hervorragende wissenschaftliche Betreuung danke ich Prof. Andreas Lübbert und
Prof. Rimvydas Simutis. Die überaus fruchtbaren nächtlichen Brainstorming-Sitzungen
bleiben unvergessen.
Ich bedanke mich insbesondere bei Herrn Prof. Lübbert für das entgegengebrachte Vertrauen
bei der Übertragung wichtiger Industrieprojekte und bei der Betreuung von Diplomanden,
sowie für die Freiheiten, die ich genoss und die unzählbaren Kongresse, die ich besuchen
durfte.
Danken möchte ich Martina Anwand und Sylke Gutknecht, ohne deren Hilfe bei der
Versuchsvorbereitung und Analytik, die große Anzahl an Experimenten alleine gar nicht zu
bewältigen gewesen wären.
Ich danke Christian Sieblist, der immer ein verlässlicher Anlaufpunkt bei Software- und
Computerfragen war. Obwohl er selbst meist knietief in eigener Arbeit steckte, hat er die Zeit
für meine Probleme entbehren können.
Des Weiteren danke ich unserem Werkstattmeister Frank Ullmann, der all meine Ideen in
praktischer Form professionell und mit einem Höchstmaß an Präzision und Funktionalität
umsetzte.
Ich danke Frau Koch und Frau Homolya für die Motivationshilfen, die sie mir in Form von
Schokolade und Gummitierchen immer wieder erfolgreich zuführten.
Bedanken möchte ich mich bei all den Projektstudenten und Diplomanden, die ich betreuen
durfte und deren Projekte einen wesentlichen Teil zu dieser Arbeit beitrugen. Deshalb vielen
Dank, Christan Berger, Silke Dietzmann, Stefan Gnoth, Markus Lange, Régis Lelou, Max
Wellerdiek, Martin Werthschulte und allen anderen, die sich angesprochen fühlen.
Ich danke meinen Eltern für die finanzielle Unterstützung während des Studiums und für die
nie versiegende Pipeline zum Kühlschrank.
Zuletzt möchte ich der Person danken, die es immer wieder geschafft hat, mit viel Liebe,
leckerem Essen und einem offenen Ohr meine Gedanken zu ordnen. Danke Kätchen!
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Contents


Chapter page

1 Introduction 1 - 15

2 Open Loop Control of the Biomass Concentration 16 - 35
within the Growth Phase of Recombinant Protein
Production Processes

3 Estimation of Biomass Concentrations in Fermentation 36 - 50
Processes for Recombinant Protein Production

4 Generic Model Control of the Specific Growth Rate in 51 - 68
Recombinant Escherichia coli Cultivations

5 Application of Model Predictive Control to Cultivation 69 - 80
Processes for Protein Production with Genetically
Modified Bacteria

6 Improving the Batch-to-Batch Reproducibility in 81 - 93
Microbial Cultures During Recombinant Protein
Production by Guiding the Process Along a Predefined
Total Biomass Profile

7 Improving the Batch-to-Batch Reproducibility of 94 - 112
Microbial Cultures During Recombinant Protein
Production by Regulation of the Total Carbon Dioxide
Production

Summary 113 - 114
Zusammenfassung 115 - 116
Publications List 117 - 119
Curriculum Vitae 120
Erklärung 121

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Chapter 1. Introduction
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Chapter 1





Introduction











Abstract. Compared to the immense achievements in fundamental molecular biological
sciences, the improvements in the fermentation and downstream processing technologies used
in industry have been less spectacular over the last decade. Hence, there is a misbalance
between new cellular systems and production technologies, resulting in a decreasing annual
rate of approvals for protein manufacturing processes. In its process analytical technology
(PAT) initiative, the U.S. Food and Drug Administration identified the issues that must be
improved to compensate for this development and forces manufactures towards a more
scientific approach of solving the problems. In this doctoral dissertation, methods of
bioprocess engineering science have been used to meet the demands. Recombinant protein
production processes, where Escherichia coli were used as host cells, are taken as a concrete
example. Concretely, a design strategy for improved robust process operational procedures
was developed that can be tightly supervised and automatically controlled.











Parts of this chapter have been published in Engineering in Life Sciences:
Jenzsch, M., Simutis, R., Lübbert, A. (2006), Optimization and Control of Industrial Microbial Cultivation
Processes, Eng. Life Sci., 6(2), 117-124
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1 Chapter 1. Introduction
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1 INTRODUCTION
In most industrial countries, biotechnology is a well recognized key technology for the years
to come. Its products or services are applied in medical care, in food technology, in
agriculture, and, increasingly, in fine chemistry. Currently most hopes are put into new
pharmaceutical products, preferentially recombinant therapeutic proteins that allow curing
diseases by directly combating the sources rather than only symptoms. Hence, much money
has been investigated into the development of new biologics.
In contrast to the immense achievements in fundamental molecular biological sciences, the
fermentation and downstream processing technologies used in industry have not been
developed at the same pace. Since they did not receive as much public interest as the
biological sciences, much less money and efforts flew into that domain. Hence, a misbalance
between new cellular systems and production technologies appeared. This resulted in a
decreasing annual rate of approved production processes over the last ten years (FDA 2005).
This development was well recognized and made public by the Food and Drug
Administration (FDA) of the USA and essentially the same observation have been made by
the European Medicines Agency (EMEA). As FDA is controlling most biologics production
processes world wide, it has the best possibilities to judge about the state of the developments.
The administration found that there are severe deficits in the production of pharmaceutical
products. Consequently the agency is forcing manufacturers to pay more attention to the
quality of their production processes. FDA does not only express its concern about stagnation
in process development, it also made suggestions to the manufactures, e.g., to make use of
recent developments in process supervision and control engineering sciences (FDA 2003). In
its “process analytical technology (PAT)” initiative, FDA proposes concretely: (i) to make use
of new measurement techniques for online supervision of the processes and (ii) to more
exhaustively exploit the measurement data gathered from the processes to gain mechanistic
understanding, predominantly knowledge about the interrelationships between the various
process variables. According to the FDA-initiative, measurement data should (iii) be used
online in order to recognize deviations from ‘in control situations’ before the processes run
out of control. And (iv) in the case of significant deviations from the desired setpoint profiles,
the processes must be drawn back to the predefined path by automatic feedback control in the
engineering sense.
One of the most