With its detailed description of membrane protein expression, high-throughput and genomic-scale expression studies, both on the analytical and the preparative scale, this book covers the latest advances in the field. The step-by-step protocols and practical examples given for each method constitute practical advice for beginners and experts alike.

Following its inception in the 1950s, cell-free protein synthesis made a tremendous impact on the basic life sciences. The use of cell-free systems was key to understanding molecular mechanisms underlying one of the most complicated processes found in nature: protein translation. Since this time, aggressive cutting-edge research and stiff commerica

This is a unique book that describes the most recent achievements in the methodology of protein biosynthesis under cell-free conditions. Various versions of cell-free protein-synthesizing systems and their applications to production of individual proteins on a preparative scale are reviewed. The most recent, advanced methodologies, such as continuous-exchange and continuous-flow cell-free systems and novel effecting batch-format cell-free procedures, are considered. Special attention is drawn to the possibilities of structural (NMR; X-ray) analysis of various gene expression products with the use of a new generation of cell-free systems.

The Nobel Prize in Medicine 1968 for interpretation of the genetic code and its function in protein synthesis and in Chemistry 2009 for studies of the structure and function of the ribosome highlighted the ground-breaking experiment performed on May 15, 1961 by Nirenberg and Matthaei and their principal breakthrough on the creation of "cell-free protein synthesis (CFPS) system". Since then the continuous technical advances have revitalized CFPS system as a simple and powerful technology platform for industrial and high-throughput protein production. CFPS yields exceed grams protein per liter reaction volume and offer several advantages including the ability to easily manipulate the reaction components and conditions to favor protein synthesis, decreased sensitivity to product toxicity, batch reactions last for multiple hours, costs have been reduced orders of magnitude, and suitability for miniaturization and high-throughput applications. With these advantages, there is continuous increasing interest in CFPS system among biotechnologists, molecular biologists and medical or pharmacologists.

During the past decade as the data on gene sequences and expression patterns rapidly accumulated, cell-free protein synthesis technology has also experienced a revolution, becoming a powerful tool for the preparation of proteins for their functional and structural analysis. In Cell-Free Protein Production: Methods and Protocols, experts in the field contribute detailed techniques, the uses of which expand deep into the studies of biochemistry, molecular biology, and biotechnology. Beginning briefly with basic methods and historical aspects, the book continues with thorough coverage of protein preparation methods, the preparation of proteins that are generally difficult to prepare in their functional forms, applications of the cell-free technologies to protein engineering, as well as some methods that are expected to constitute a part of future technologies. Written in the highly successful Methods in Molecular BiologyTM series format, the chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Cell-Free Protein Production: Methods and Protocols aims to help researchers continue the growth of the vital exploration of cell-free sciences and technologies in order to better understand the dynamic lives of cells.

Cell-free synthetic biology is in the spotlight as a powerful and rapid approach to characterize and engineer natural biological systems. The open nature of cell-free platforms brings an unprecedented level of control and freedom for design compared to in vivo systems. This versatile engineering toolkit is used for debugging biological networks, constructing artificial cells, screening protein library, prototyping genetic circuits, developing new drugs, producing metabolites, and synthesizing complex proteins including therapeutic proteins, toxic proteins, and novel proteins containing non-standard (unnatural) amino acids. The book consists of a series of reviews, protocols, benchmarks, and research articles describing the current development and applications of cell-free synthetic biology in diverse areas.

This book reviews cell-free production systems, exploring the frontiers in cellular engineering and biotechnology. With contributions from experts in the field, the book offers a comprehensive and up-to-date account of the latest advancements and practical applications. The volume covers a diverse range of topics, beginning with an in-depth analysis of cell-free display techniques for protein evolution, shedding light on the methodologies used to engineer proteins for diverse purposes, followed by an examination of bottom-up synthetic biology employing cell-free protein synthesis. Additionally, it investigates the intricacies of the cell-free synthesis of metalloproteins, elucidating the unique properties and functionalities of these biologically important molecules. In this book, particular attention is given to the integration of cell-free production systems with droplet microfluidics, a pioneering approach that has revolutionized research activities in both academic and industrial settings. Readers will also discover the latest advancements in cell-free protein synthesis and immobilization, and find out more about the eCell technology, which combines cell-free protein synthesis with bio-sensing and remediation, revolutionizing critical areas of study in biotechnology. Together with the companion volume entitled “Cell-free Production: System Development”, both books highlight the research progresses on the basic and applied research of cell-free production systems in the last few years, and are invaluable resources for scholars, researchers, and bioengineers. This book also appeals to enthusiasts of synthetic biology.

This book describes advanced studies in cell-free synthetic biology, an emerging biotechnology that focuses on cell-free protein synthesis and cell-free systems for fundamental and industrial research in areas such as genetic circuit design, small-molecule synthesis, complicated-macromolecule synthesis, unnatural-macromolecule synthesis, high-throughput screening, artificial cells, and biomaterials. Cell-free synthetic biology is now an integral part of developing fields like nanotechnology, materials science, and personalized medicine. The book discusses the main research directions in the development of cell-free systems, as well as a number of applications of cell-free synthetic biology, ranging from structural biology to the human health industry. It is intended for students and researchers in life sciences, synthetic biology, bioengineering, and chemical engineering.