Recombinant Protein Production in Yeast
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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. Front Matter ....Pages i-xii Established and Upcoming Yeast Expression Systems (Burcu Gündüz Ergün, Damla Hüccetoğulları, Sibel Öztürk, Eda Çelik, Pınar Çalık)....Pages 1-74 Detection and Elimination of Cellular Bottlenecks in Protein-Producing Yeasts (Richard J. Zahrl, Brigitte Gasser, Diethard Mattanovich, Pau Ferrer)....Pages 75-95 Pichia pastoris Promoters (Aysun Türkanoğlu Özçelik, Semiramis Yılmaz, Mehmet Inan)....Pages 97-112 Selection of the Optimal Yeast Host for the Synthesis of Recombinant Enzymes (Felix Bischoff, Martin Giersberg, Falko Matthes, Tobias Schwalenberg, Sebastian Worch, Gotthard Kunze)....Pages 113-132 Screening and Selection of Production Strains: Secretory Protein Expression and Analysis in Hansenula polymorpha (Yoo Su Jin, Moon Hye Yun, Kang Hyun Ah)....Pages 133-151 Selection of Heterologous Protein-Producing Strains in Yarrowia lipolytica (Paul Soudier, Macarena Larroude, Ewelina Celińska, Tristan Rossignol, Jean-Marc Nicaud)....Pages 153-168 High-Throughput Screening and Selection of Pichia pastoris Strains (Roland Weis)....Pages 169-185 Analyzing Recombinant Protein Production in Pichia pastoris with Targeted Proteomics (Roslyn M. Bill, Annegret Ulke-Lemée, Stephanie P. Cartwright, Rena Far, Jay Kim, Justin A. MacDonald)....Pages 187-209 CRISPR/Cas9-Mediated Homology-Directed Genome Editing in Pichia pastoris (Thomas Gassler, Lina Heistinger, Diethard Mattanovich, Brigitte Gasser, Roland Prielhofer)....Pages 211-225 Tailoring N-Glycan Biosynthesis for Production of Therapeutic Proteins in Saccharomyces cerevisiae (Mari A. Piirainen, Heidi Salminen, Alexander D. Frey)....Pages 227-241 Improving the Production of Cofactor-Containing Proteins: Production of Human Hemoglobin in Yeast (Olena P. Ishchuk, José L. Martínez, Dina Petranovic)....Pages 243-264 Membrane Protein Production in Yeast: Modification of Yeast Membranes for Human Membrane Protein Production (Anita Emmerstorfer-Augustin, Tamara Wriessnegger, Melanie Hirz, Guenther Zellnig, Harald Pichler)....Pages 265-285 Yeast Surface Display and Cell Sorting of Antigen-Binding Fc Fragments (Flávio Sádio, Gerhard Stadlmayr, Katharina Stadlbauer, Florian Rüker, Gordana Wozniak-Knopp)....Pages 287-308 Guidelines for Small-Scale Production and Purification of Hepatitis B Surface Antigen Virus-Like Particles from Recombinant Pichia pastoris (Maria Zahid, Ursula Rinas)....Pages 309-322 Efficient Development of a Mixed Feed Process for Pichia pastoris (David Johannes Wurm, Oliver Spadiut)....Pages 323-333 Primary Recovery of Yeast Culture Supernatant for Recombinant Protein Purification (Michael M. Maurer, Harald Schillinger)....Pages 335-342 Purification of Recombinant Glycoproteins from Pichia pastoris Culture Supernatants (David Johannes Wurm, Oliver Spadiut)....Pages 343-350 LC-MS Analysis of (Glyco-)Proteins of Pichia pastoris (Clemens Grünwald-Gruber, Friedrich Altmann)....Pages 351-360 Assessing the Quality of Recombinant Products Made in Yeast (Karola Vorauer-Uhl, Gabriele Lhota)....Pages 361-384 Quantification of Recombinant Products in Yeast (Karola Vorauer-Uhl, Gabriele Lhota)....Pages 385-428 Back Matter ....Pages 429-431
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