This book provides a detailed and up-to-date overview of all aspects of yeast cell surface engineering, including fundamental principles, practical strategies for the construction of engineered yeasts, as well as medical and industrial applications. The technique makes it possible to add eukaryotic modifications to the surface-displayed proteins/peptides, which is of significant value in basic and applied research. Generally referred to as an arming (molecular display) technology, it allows yeast to be used as a whole-cell biocatalyst for a range of purposes, including bio-energy production, pollutant removal, recovery of rare metal ions, and preparation of functional cells, all of which are comprehensively covered in the book. Among the medical applications discussed are in vitro antibody preparation and the production of oral vaccines. In addition, it presents the latest advances in protein engineering and high-throughput screening for directed evolution of enzymes. The book enables graduate students and researchers to gain a deeper, comprehensive understanding of the technology, and offers further inspiration for researchers and industrial experts in this rapidly evolving field.

A summary of the recent achievements in surface-functionalised cells including fabrication, characterisation, applications and nanotoxicity.

In addition to research and discovery, yeast surface display technology has found applications in industrial processes such as biofuel production and environmental pollutant absorption and degradation. Yeast Surface Display: Methods, Protocols, and Applications guides readers through yeast surface antibody display library and antibody engineering, yeast surface display as a tool for protein engineering, yeast surface cDNA display library construction and applications, and yeast surface display in bioassay and industrial applications. 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 key tips on troubleshooting and avoiding known pitfalls. Concise and easy-to-use, Yeast Surface Display: Methods, Protocols, and Applications aims to help accelerate the work of protein chemists, antibody engineers, molecular and cell biologists, and industrial bioengineers. ​

This detailed volume explores a wide variety of applications of yeast surface display, an extensively used protein engineering technology. Beginning with detailed protocols for the construction and efficient selection/screening of yeast surface display libraries, as well as for the analysis of individual yeast-displayed protein variants, the book continues with protocols describing the selection of yeast surface display libraries for binding to mammalian cells or to extracellular matrix as well as protocols for a broad spectrum of specialized yeast surface display applications, demonstrating the versatility of this display platform. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible methodologies, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Yeast Surface Display serves as a comprehensive resource that enables the implementation of this powerful and versatile technique in virtually any molecular biology laboratory, even in the absence of any prior yeast surface display experience.

Cell surface engineering is an emerging field concerning cell surface modifications to enhance its functionalities. The book introduces the reader to the area of surface-functionalized cells and summarizes recent developments in the area including fabrication, characterization, applications and nanotoxicity. Topics covered include recent approaches for the functionalization of cells with nanomaterials (polymer nanofilms and nanoparticles), fabrication of functional biomimetic devices and assemblies based on nanoparticle-modified microbial cells and artificial spores (the bioinspired encapsulation of living cells with inorganic nanoshells). The book provides an interdisciplinary approach to the topic with authors from both biological and chemical backgrounds. This multidisciplinary view makes the book suitable for those interested in biomaterials, biochemistry, microbiology and colloid chemistry, providing both an introduction for postgraduate students as well as a comprehensive summary for those already working in the area. biomaterials, biochemistry, microbiology and colloid chemistry.

In addition to research and discovery, yeast surface display technology has found applications in industrial processes such as biofuel production and environmental pollutant absorption and degradation. Yeast Surface Display: Methods, Protocols, and Applications guides readers through yeast surface antibody display library and antibody engineering, yeast surface display as a tool for protein engineering, yeast surface cDNA display library construction and applications, and yeast surface display in bioassay and industrial applications. 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 key tips on troubleshooting and avoiding known pitfalls. Concise and easy-to-use, Yeast Surface Display: Methods, Protocols, and Applications aims to help accelerate the work of protein chemists, antibody engineers, molecular and cell biologists, and industrial bioengineers. ​

Enzyme engineering is a valuable tool for generating powerful and robust biocatalysts for a broad range of applications. Directed evolution using cell surface display has been previously applied for enzyme engineering, and involves iterative optimization of a protein sequence to yield a desired phenotype. However, previous applications have been carefully tailored for specific enzymes, and this process of developing a good screen is a significant bottleneck in engineering. Co-display of an enzyme and its substrate on the yeast surface is a promising general strategy for evolving bond-forming enzymes, but has only been used to engineer one enzyme. We extended yeast surface co-display to engineer Sfp, a bioconjugation enzyme, for improved activity with a non-native substrate. Sfp library screening identified mutational hotspots in the enzyme and led to the design of Sfp variants with orders of magnitude enhancements in activity. However, these activity improvements were only apparent in contexts similar to yeast surface co-display, reflecting artifacts in the use of this approach for enzyme engineering. Based on our results, we offer suggestions for improving the success of engineering strategies based on yeast surface display.