Institut für Verfahrenstechnik, Umwelttechnik und Techn. Biowissenschaften
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Method establishment for protein expression quantification

 

Wir bitten um Verständnis, dass die Informationen über dieses Projekt derzeit nur in englischer Sprache verfügbar sind.

 

Project Content:

Recombinant protein production is the most promising approach to produce new products for tailored pharmaceutical therapies. The main goal of this project is to establish a methodology to characterize a recombinant strain regarding the protein production process by means of increased process understanding, hence, to identify the cell state respective to recombinant production in real-time by direct or indirect methods. Particularly, we aim to characterize the methanol-inducible protein production in a recombinant Pichia pastoris strain. 
Fast and easy to handle experiments were carried out, using batch and fed-batch cultures in highly instrumented bioreactor configurations, including various non-invasive, in-line and at-line sensors, coupled to a process management system, which allows deriving information from data by real-time calculation functionality. 
Using dynamic culture conditions in batch experiments critical parameters as the specific methanol uptake rate, qs, was determined for different strains. Based on specific uptake rates, several feeding strategies were developed. It could be demonstrated that a faster increase of specific substrate uptake rate, and heading off from maintenance effects, leads to higher specific productivity. 
Future work will focus on mixed feed production strategies to optimize recombinant protein production based on specific uptake rates. Additional, a focus will be on direct measurement of a product with characteristic growth and production related parameters. Those parameters represent a generic approach to know the production process in real time.


 

Scientific contributions:

Thesis:

Dietzsch C. 2012. Protein expression in Pichia pastoris - A novel strategy for fast bioprocess development

 

Publications:

Zalai D, Dietzsch C, Herwig C, Spadiut O. 2012. A dynamic fed batch strategy for a Pichia pastoris mixed feed system to increase process understanding. Biotechnol Prog. doi: 10.1002/btpr.1551

Krainer F, Dietzsch C, Hajek T, Herwig C, Spadiut O, Glieder A. 2012. Recombinant protein expression in Pichia pastoris strains with an engineered methanol utilization pathway. Microbial Cell Factories 11(1):22.

Dietzsch C, Spadiut O, Herwig C. 2011. A dynamic method based on the specific substrate uptake rate to set up a feeding strategy for Pichia pastoris. Microb. Cell Fact. 10(1):14.

Dietzsch C., Spadiut, O., Herwig, C. 2011. A fast approach to determine a fed batch feeding profile for recombinant Pichia pastoris strains. Microb. Cell Fact. 10, (1), 85.

 

Talks:

Spadiut O., Dietzsch C., Krainer F., Hajek T., Glieder A.,Herwig C.: "Playing with the methanol utilization pathwy in Pichia pastoris";Pichia 2012 Conference, Alpbach, Asutria; 29.02.2012-03.03.2012.

C. Dietzsch, C. Herwig:"Bioprozessoptimierung eines rekombinanten Pichia pastoris-Expressionsstammes";
Vortrag: ProcessNet Jahrestagung 2010, Aachen, Deutschland; 21.09.2010 - 23.09.2010; in: "Chemie Ingenieur Technik", Chemie Ingenieur Technik / Wiley-VCH, 82 / 9 (2010), ISSN: 0009-286x; S. 1494 - 1495.

C. Dietzsch, C. Herwig: "Determination of protein production related parameters in a recombinant Pichia Pastoris MutS expression strain";
Vortrag: ESBES - 8th European Symposium on Biochemical Engineering Science, Bologna, Italien; 05.09.2010 - 08.09.2010; in: "Book of Abstracts", (2010), S. 155.

 

Poster:

C.Dietzsch, O. Spadiut, C. Herwig: "From Strain to Product: A fast approach in bioprocess development for recombinant protein expression in Pichia pastoris"; Pichia 2012 Conference, Alpbach, Asutria; 29.02.2012-03.03.2012.

C. Dietzsch, O. Spadiut, C. Herwig; “A dynamic method based on the specific substrate uptake rate to set up a feeding strategy for Pichia Pastoris“; BioProduction Forum 2011; Ulm; Germnay; 08.06.2011-10.06.2011, 1st Poster award.

C. Dietzsch, O. Spadiut, C. Herwig; “A dynamic method based on the specific substrate uptake rate to set up a feeding strategy for Pichia Pastoris“; 6th Conference on recombinant protein production; Vienna; Austria; 16.02.2011-19.02.2011.

C. Dietzsch, D. Rial, C. Herwig: "Method establishment for determination of protein expression related parameters"; International Graz Congress for Pharmaceutical Engineering, Graz; Austria; 17.09.2009 - 18.09.2009; in: "International Graz Congress for Pharmaceutical Engineering - Book of Abstracts", (2009).

D. Rial, C. Dietzsch, C. Herwig: "Comprehensive characterization of recombinant Pichia pastoris using dynamic process conditions and process analytical technology"; 14th European Congress on Biotechnology, Barcelona, Spain; 13.09.2009 - 16.09.2009; in: "Abstracts of the 14th European Congress on Biotechnology", New Biotechnology / Elsevier, 25 (Supplement) (2009), ISSN: 1871-6784; S. 204 - 205.

 

Kontakt:

Univ.Prof. Dipl.-Ing. Dr.techn. Christoph Herwig

Mitarbeiter:

Dipl.-Ing. Dr.techn. Christian Dietzsch

Univ.Ass. Dipl.-Ing. Dr.nat.techn. Oliver Spadiut

Externe Partner:

Prof. Anton Glieder; TU Graz; Austria