2.Detection and Elimination of Cellular Bottlenecks in Protein Producing Yeasts
Richard J. Zahrl, Brigitte Gasser, Diethard Mattanovich, and Pau Ferrer
3.Pichia pastoris Promoters
Aysun Özçelik, Semiramis Yılmaz, and Mehmet Inan
4. Selection of the Optimal Yeast Host for the Synthesis Recombinant Enzymes
Felix Bischoff, Martin Giersberg, Falko Matthes, Tobias Schwalenberg, Sebastian Worch, and Gotthard Kunze
5. Screening and Selection of Production Strains: Secretory Protein Expression and Analysis in Hansenula polymorpha
Su Jin Yoo, Hye Yun Moon, and Hyun Ah Kang
6. Selection of Heterologous Protein Producing Strains in Yarrowia lipolytica
Paul Soudier, Macarena Larroude, Ewelina Celinska, Tristan Rossignol, and Jean-Marc Nicaud
7. High-throughput Screening and Selection of Pichia pastoris Strains
Roland Weis
8. Analyzing Recombinant Protein Production in Pichia pastoris with Targeted Proteomics Roslyn M. Bill, Annegret Ulke-Lemée, Stephanie P. Cartwright, Rena Far, Jay Kim, and Justin A. MacDonald
9. CRISPR/Cas9-mediated Homology Directed Genome Editing in Pichia pastori
Thomas Gassler, Lina Heistinger, Diethard Mattanovich, Brigitte Gasser, and Roland Prielhofer
10. Tailoring N-glycan Biosynthesis for Production of Therapeutic Proteins in Saccharomyces cerevisiae
Mari A. Piirainen, Heidi Salminen and Alexander D. Frey
11. Improving the Production of Cofactor Containing Proteins: Production of Human Hemoglobin in Yeast
Olena P. Ishchuk, José L. Martínez, and Dina Petranovic
12. Membrane Protein Production in Yeast: Modification of Yeast Membranes for Human Membrane Protein Production
Anita Emmerstorfer-Augustin, Tamara Wriessnegger, Melanie Hirz, Guenther Zellnig, and Harald Pichler
13. Yeast Surface Display and Cell Sorting of Antigen-binding Fc Fragments
Flávio Sádio, Gerhard Stadlmayr, Katharina Stadlbauer, Florian Rüker, and Gordana Wozniak-Knopp
14. Guidelines for Small-scale Production and Purification of Hepatitis B surface Antigen Virus-like Particles from Recombinant Pichia pastoris
Maria Zahid and Ursula Rinas
15. Efficient Development of a Mixed Feed Process for Pichia pastoris
David Johannes Wurm and Oliver Spadiut
16. Primary Recovery of Yeast Culture Supernatant for Recombinant Protein Purification
Michael M. Maurer and Harald Schillinger
17. Purification of Recombinant Glyco-proteins from Pichia pastoris Culture Supernatants
David Johannes Wurm and Oliver Spadiut
18. LC-MS analysis of (glyco)-proteins of Pichia pastoris
Clemens Grünwald-Gruber and Friedrich Altmann
19. Assessing the Quality of Recombinant Products Made in Yeast
Karola Vorauer-Uhl and Gabriele Lhota
20. Quantification of Recombinant Products In Yeast
Karola Vorauer-Uhl and Gabriele Lhota
This volume provides an overview of the main yeast production platforms currently used and future yeast cell factories for recombinant protein production. Chapters detail approaches of genetic and metabolic engineering, co-factor containing proteins and virus-like particles, glycoproteins, and post-translational modifications of proteins. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Recombinant Protein Production in Yeast: Methods and Protocols aims to provide state of the art background and methods for protein producing yeast platforms, as well as case studies for special applications.