This book covers the fundamentals of protein inactivation during bioseparation and the effect on protein processing. Bioseparation of Proteins is unique because it provides a background of the bioseparation processes, and it is the first book available to emphasize the influence of the different bioseparation processes on protein inactivation. Bioseparation of Proteins covers the extent, mechanisms of, and control of protein inactivation during these processes along with the subsequent and essential validation of these processes. The book focuses on the avoidance of protein (biologicalproduct) inactivation at each step in a bioprocess. It compares protein inactivation exhibited during the different bioseparation processes by different workers and provides a valuable framework for workers in different areas interested in bioseparations. Topics include separation and detection methods; estimates of protein inactivation and an analysis of this problem for different separation processes; strategies for avoiding inactivation; the molecular basis of surface activity and protein adsorption,process monitoring, and product validation techniques; and the economics of various bioseparation processes and quality control procedures. Key Features * Protein inactivation and other aspects of biological stability are critical to an effective bioseparation process; This book is a detailed and critical review of the available literature in an area that is essential to the effectiveness, validation, and economics of bioseparation processes for drugs and other biological products; Conveniently assembled under one cover, the survey of the literature and resulting perspective will greatly assist engineers and chemists in designingand improving their own processes; Key features of the text include: * detailed data on biological stability under various bioseparation conditions * extensive case studies from the literature on separation processes, validation, and economics * simplified analysis of protein refolding and inactivation mechanisms * consideration of adsorption theories and the effect of heterogeneity * coverage of both classical and novel bioseparation techniques, including chromatographic procedures

The biopharmaceutical industry has become an increasingly important player in the global economy, and the success of these products depends on the development and implementation of cost-effective, robust and scaleable production processes. Bioseparations-also called downstream processing- can be a key source of competitive advantageto biopharmaceut

Rapid advances in molecular biology have accelerated the production of a great number of protein-based therapeutic agents. The major cost in producing these proteins appears to be associated with their purification from the complex mixture of the crude extract. A major challenge to the protein biochemist and the biochemical engineer is the development of rapid, efficient, and cost-effective purification systems. This volume presents state-of-the-art reviews of current methods used in the purifica tion of biological macromolecules that are based on molecular interactions. Thus, the major emphasis is placed on affinity-related techniques. Part I provides a general introduction to affinity chromatography and includes a chapter describing an interesting new technique called "slalom chromatography" for DNA fractionations. Affinity chromatography using molecules of biological origin as the affinity ligand is covered in Part II. Part III describes the use of a special class of biomolecules, antibodies, as affinity ligands. Affinity chromatography with biomimetic ligands is discussed in Part IV. Newer concepts and their applications in bioseparation are presented in Part V. Part VI covers affinity-related techniques such as affinity-based extracorporeal shunts, affinity electrophoresis, affinity precipitation, and affinity extraction. I would like to express my sincere thanks to all the authors, who are recognized experts in their respective fields, for their cooperation and contributions. I thank the editorial staff of Plenum Press for their professionalism, and Mary Phillips Born, Senior Editor, for her encouragement. The support of my family (Ping and Peilin) made it possible to complete editing this book. That T.

This publication details the isolation of proteins from biological materials, techniques for solid-liquid separation, concentration, crystallization, chromatography, scale-up, process monitoring, product formulation, and regulatory and commercial considerations in protein production. The authors discuss the release of protein from a biological host, selectivity in affinity chromatography, precipitation of proteins (both non-specific and specific), extraction for rapid protein isolation, adsorption as an initial step for the capture of proteins, scale-up and commercial production of recombinant proteins, and process monitoring in downstream processing.

Ultrafiltration (UF) is a pressure-driven separation process in which membranes are used for a broad variety of applications, ranging from the processing of biological macromolecules to wastewater treatment. It has significant advantages over competing separation technologies. Food and biotechnological applications of UF account for nearly 40 percent of the current total usage. In the case of high value therapeutic protein and DNA products, the separation and purification costs can be as high as 80 percent of the total cost of production. Therefore, it makes economic sense to develop cost-effective and scaleable purification processes for such products. UF is used for protein concentration, protein desalting and protein fractionation (such as protein-protein separation). Concentration and desalting processes are technologically less demanding and have been widely used in the bioprocess industry for quite some time. Protein fractionation, on the other hand, is a challenging proposition and is definitely a more recent development. This text focuses primarily on protein fractionation.

The challenge of bioseparations is to isolate and purify identified products from the dilute product broth produced from cell culture. Innovation in bioseparations technology is increasingly driven by the requirements imposed by the growing importance of production on a process scale of injectable-grade products, and economic pressures to improve the efficiency of downstream processing. As in other areas of technical change, science does not necessarily precede new technology: progress results from a complex and messy mixture of advances in understanding, ingenious ideas, novel techniques and chance discoveries. What is certain is that close interaction between academics and practitioners, biological scientists and process engineers is needed to solve the problems of bioseparations. The Second International Conference on Separations for Biotechnology at Reading, UK, in September 1990 set out to provide a critical multidisciplinary forum for the discussion of bioseparations. This volume contains the papers presented at the meeting. The meeting was organised around six themes with oral and poster presentations on the science and practice of bioseparations technology, and the same structure has been kept for this book. We have also included the texts of the keynote review paper by Professor Alan Michaels and the introductory review papers specially commissioned for the conference. Within each part of this book the review paper is followed by the contributed papers grouped alphabetically by their first author. All the original papers published here were accepted for publication after scientific refereeing.

It is generally recognized that the commercial success of biotechnology products is highly dependent on the successful development and application of high-powered separation and purification methods. In this practical and authoritative handbook, the separation of proteins, nucleic acids, and oligonucleotides from biological matrices is covered from analytical to process scales. Also included in a chapter on the separation of monoclonal antibodies, which have found numerous uses as therapeutic and diagnostic agents. Analytical techniques include an interesting montage of chromatographic methods, capillary electrophoresis, isoelectric focusing, and mass spectrometry. Among separation and purification methods, liquid-liquid distribution, displacement chromatography, expanded bed adsorption, membrane chromatography, and simulated moving bed chromatography are covered at length. Regulatory and economic considerations are addressed, as are plant and process equipment and engineering process control. A chapter on future developments highlights the application of DNA chip arrays as well as evolving methodologies for a large number of drugs that are under development for treatment of cancer, AIDS, rheumatoid arthritis, and Alzheimer's disease. Handbook of Bioseparations serves as an essential reference and guidebook for separation scientists working in the pharmaceutical and biotechnology industries, academia, and government laboratories. Key Features * Covers bioseparations of proteins, nucleic acids, and monoclonal antibodies * Encompasses both analytical and process-scale methods * Elucidates the importance of engineering process control * Details selection of plant and process equipment * Addresses economic considerations * Discusses future developments