Dynamic modeling of methylotrophic Pichia pastoris culture with exhaust gas analysis: From cellular metabolism to process simulation

Niu, Hongxing; Daukandt, Marc; Rodriguez, Christian; Fickers, Patrick; Bogaerts, Philippe

doi:10.1016/j.ces.2012.11.006

Cited by 17 publications

(18 citation statements)

References 33 publications

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“…Volumetric rates,

r_{{normalO}_{2}}

and

r_{{normalCO}_{2}}

would provide direct information about the growth rate and their ratio, thus indirectly implying cellular respirative/fermentative metabolism (Niu et al . ) of P. pastoris . Therefore, OUR and CER measurements were also recorded in the SCADA, and a dynamic plot of OUR was generated against time.…”

Section: Methodsmentioning

confidence: 99%

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High level extracellular production of recombinant human interferon alpha 2b in glycoengineeredPichia pastoris:culture medium optimization, high cell density cultivation and biological characterization

et al. 2019

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Aims The present study was aimed at design of experiments (DoE)‐ and artificial intelligence‐based culture medium optimization for high level extracellular production of a novel recombinant human interferon alpha 2b (huIFNα2b) in glycoengineered Pichia pastoris and its characterization. Methods and Results The artificial neural network—genetic algorithm model exhibited improved huIFNα2b production and better predictability compared to response surface methodology. The optimized medium exhibited a fivefold increase in huIFNα2b titre compared to the complex medium. A maximum titre of huIFNα2b (436 mg l−1) was achieved using the optimized medium in the bioreactor. Real‐time capacitance data from dielectric spectroscopy were utilized to model the growth kinetics with unstructured models. Biological characterization by antiproliferative assay proved that the purified recombinant huIFNα2b was biologically active, exhibiting growth inhibition on breast cancer cell line. Conclusions Culture medium optimization resulted in enhanced production of huIFNα2b in glycoengineered P. pastoris at both shake flask and bioreactor level. The purified huIFNα2b was found to be N‐glycosylated and biologically active. Significance and Impact of the Study DoE‐based medium optimization strategy significantly improved huIFNα2b production. The antiproliferative activity of huIFNα2b substantiates its potential scope for application in cancer therapy.

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“…Volumetric rates,

r_{{normalO}_{2}}

and

r_{{normalCO}_{2}}

Section: Methodsmentioning

confidence: 99%

“…Apart from dielectric spectroscopy, exhaust gas analysis or respirometry has also been used as an online tool for biomass monitoring by quantifying oxygen uptake rate (OUR) and carbon dioxide evolution rate (CER; Niu et al . ).…”

Section: Introductionmentioning

confidence: 97%

High level extracellular production of recombinant human interferon alpha 2b in glycoengineeredPichia pastoris:culture medium optimization, high cell density cultivation and biological characterization

et al. 2019

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“…Volumetric rates,

r_{O_{2}} and r_{{CO}_{2}}

could provide direct insight on the cell physiology and their ratio indicates the respiratory quotient of Pichia pastoris . OUR and CER values were estimated by SCADA and logged at every 10 s interval.…”

Section: Methodsmentioning

confidence: 99%

Control of specific growth rate for the enhanced production of human interferon α2b in glycoengineered Pichia pastoris: process analytical technology guided approach

Katla

Mohan

Pavan

et al. 2019

J of Chemical Tech & Biotech

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BACKGROUND Process variability in bioprocess systems involving genetically engineered strains is a common bottleneck and a real‐time insight of the on‐going process is crucial to achieve the desired product and its quality. In this study, a process analytical technology (PAT) platform was developed for the monitoring and control of specific growth rate during methanol induction phase (μmet) of glycoengineered Pichia pastoris fermentation for human interferon alpha 2b (huIFNα2b) production. RESULTS The PAT guided approach involves real‐time monitoring of capacitance (ΔC) facilitating online estimation of specific growth rate (μest), which serves as a process input during controller application. Fed‐batch experiments using pulsed‐feeding of methanol at different dosage (20 g and 30 g) did not significantly influence μmet. Exponential methanol feeding was achieved using a modified proportional, integral and derivative (PID) controller for different predefined specific growth rate set point (μsp) values, namely 0.015, 0.03, 0.04 and 0.06 h−1. Exponential feeding strategy during the induction phase resulted in two crucial outcomes: (i) controlled methanol feeding rate (regulated by the developed PID controller) balanced the methanol consumption rate (qs, met) of the P. pastoris; (ii) significant improvement in huIFNα2b titer (1483 mg L−1) and specific productivity(>0.4 mg g−1. h) was achieved by the robust control of μmet at optimal (0.04 h−1) value. The purified huIFNα2b was found to exhibit antiproliferative effect against human breast cancer cell lines. CONCLUSIONS Efficient control of μmet at a very low narrow range was achieved with a long‐term controller stability (>10 h) and the highest titer reported to date for huIFNα2b (at optimal μmet) in the yeast expression platform. © 2019 Society of Chemical Industry

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“…Considering that there are several media compounds that could be used for control, more complex models that take the underlying metabolic network into account seem to be warranted. This idea is not new and has been used for the modeling of other bioprocesses [28][29][30][31][32]. Metabolic flux networks of mammalian cells have been investigated by a number of researchers [33,34].…”

Section: Mathematical Modellingmentioning

confidence: 99%

Quality by control: Towards model predictive control of mammalian cell culture bioprocesses

Sommeregger¹,

Sissolak

Kandra

et al. 2017

Biotechnology Journal

141

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The industrial production of complex biopharmaceuticals using recombinant mammalian cell lines is still mainly built on a quality by testing approach, which is represented by fixed process conditions and extensive testing of the end-product. In 2004 the FDA launched the process analytical technology initiative, aiming to guide the industry towards advanced process monitoring and better understanding of how critical process parameters affect the critical quality attributes. Implementation of process analytical technology into the bio-production process enables moving from the quality by testing to a more flexible quality by design approach. The application of advanced sensor systems in combination with mathematical modelling techniques offers enhanced process understanding, allows on-line prediction of critical quality attributes and subsequently real-time product quality control. In this review opportunities and unsolved issues on the road to a successful quality by design and dynamic control implementation are discussed. A major focus is directed on the preconditions for the application of model predictive control for mammalian cell culture bioprocesses. Design of experiments providing information about the process dynamics upon parameter change, dynamic process models, on-line process state predictions and powerful software environments seem to be a prerequisite for quality by control realization. Keywords: Biopharmaceutical production · Design of experiments · Mathematical modelling · Process analytical technology · Soft-sensingCorrespondence: Dr. Gerald Striedner, Department of Biotechnology (DBT), University of Natural Resources and Life Sciences (BOKU), Muthgasse 18, 1190 Vienna, Austria E-mail: gerald.striedner@boku.ac.at Abbreviations: CHO, Chinese hamster ovary; CPP, critical process parameter; CQA, critical quality attribute; DCS, distributed control system; DoE, design of experiments; FDA, Food and Drug Administration; iDoE, intensified design of experiments; MBDoE, model based design of experiments; MPC, model predictive control; MVDA, multivariate data analysis; PAT, process analytical technology; QbD, quality by design; SCADA, supervisory control and data acquisition; TPP, target product profile

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Dynamic modeling of methylotrophic Pichia pastoris culture with exhaust gas analysis: From cellular metabolism to process simulation

Cited by 17 publications

References 33 publications

High level extracellular production of recombinant human interferon alpha 2b in glycoengineeredPichia pastoris:culture medium optimization, high cell density cultivation and biological characterization

High level extracellular production of recombinant human interferon alpha 2b in glycoengineeredPichia pastoris:culture medium optimization, high cell density cultivation and biological characterization

Control of specific growth rate for the enhanced production of human interferon α2b in glycoengineered Pichia pastoris: process analytical technology guided approach

Quality by control: Towards model predictive control of mammalian cell culture bioprocesses

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