High-Throughput Protein Production and Purification: Methods and Protocols(2025 Methods in Molecular Biology)

This book compiles key protocols instrumental to the study of high-throughput protein production and purification which have been refined and simplified over the years and are now ready to be transferred to any laboratory. Beginning with a section covering general procedures for high-throughput protein production, the volume continues with high-throughput protocols adapted to the production of specific protein families, as well as an extensive section on protocols combining high-throughput protein production and their micro-characterization. Written for the highly successful Methods in Molecular Biology series, chapters in this book 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 practical, High-Throughput Protein Production and Purification: Methods and Protocols serves biochemists ranging from engineers, PhD students and post-doctoral fellows, to the heads of protein expression facilities and researchers, in pursuing this vital area of study.

Table of Contents:
Part 1: General procedures for high-throughput protein production.- Overview of high-throughput cloning methods for the post-genomic era.- Overview of a high-throughput pipeline for streamlining the production of recombinant proteins.- Semi-automated small-scale purification method for high-throughput expression analysis of recombinant proteins.- High-Throughput Protein Production in Yeast.- A High-Throughput System for Transient and Stable Protein Production in Mammalian Cells.- A High-throughput Automated Protein Folding System.- Part 2: High-throughput Protocols adapted to the production of specific protein families.- High-throughput production of oxidized animal toxins in Escherichia coli.- High-Throughput purification of protein kinases in Escherichia coli and insect cells.- Parallelized micro-scale expression of soluble scFv.- High throughput production of influenza virus-like particle (VLP) array by using VLP-factoryTM, a MultiBac baculoviral genome customized for enveloped VLPexpression.- High-throughput Protein Production of membrane proteins in Saccharomyces cerevisiae.- High-throughput E. coli cell-free expression: From PCR product design to functional validation of GPCR.- High-Throughput Site-Directed Mutagenesis.- Part 3: High-throughput protein production combined with High-throughput micro-characterization for protein stability, solubility, quality and interactions.- Hot CoFi blot – A High-throughput colony-based screen for identifying more thermally stable protein variants.- High-throughput isolation of soluble protein domains using the bipartite split-GFP system.- High-throughput Analytical Light Scattering for Protein Quality Control and Characterization.- High-throughput Nano-Scale Characterization of Membrane Proteins Using Fluorescence-Detection Size-Exclusion Chromatography.- A Flexible and Scalable High-Throughput Platform for Recombinant Membrane Protein Production.- Adaption of the Leishmania cell-free expression system to high-throughput analysis of protein interactions.- High-throughput protein-protein interaction assays using tripartite split-GFP complementation.- High-throughput production of a new library of human single and tandem PDZ domains allows quantitative PDZ-peptide interaction screening through high throughput holdup assay.- High throughput protein analysis using Negative Stain Electron Microscopy and 2D classification.- High-throughput protein production combined with High throughput SELEX identifies an extensive atlas of Ciona robusta transcription factor DNA-binding specificities.- High-throughput Micro-characterization of RNA-protein interactions.