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High performance liquid chromatography in biotechnology
BOOK REVIEWS
1205
There are times also when I would have preferred a more detailed description of the underlying assumptions of a given technique, and more mathematical formulae, but these are relatively small matters. The book begins with revision of r- and F-tests, which leads naturally to simple analysis of variance techniques. Experimental design methods are then described for systems of increasing complexity. Factorial design then follows, and then fractional factorial design. The book concludes with a chapter on response surfaces. This is the right subject matter, well handled, and I recommend this book as a good introduction to chemometrics. C. J. GILMORE High Performance Liquid Chromatographyin Bioted~m-dogy:W. S. HANCOCK (editor), Wiley-Interscience, New York, 1990. Pages x + 564, E74.35. Lavoisier’s axiom that only pure substances would yield information of significance is as true today as it was in his day. We now possess the techniques to enable us to isolate well-defined compounds that satisfy the most stringent of purity. There can be no doubt that high performance liquid chromatography has proved to be one of the most important and rapidly advancing of these techniques. Since biotechnology is another branch of science which is making rapid strides and which offers considerable promise for the future, this book, which examines the various applications of HPLC to biotechnology is very timely indeed. It is worth noting that the majority of papers published recently which are concerned with HPLC have concentrated on biological applications. The book nicely reflects the multifaceted nature of the subject and the important interplay between its analytical and preparative aspects. The range of applications stretches from the nanogram scale, for characterization studies of novel proteins, to the kilogram amounts required for a manufacturing process. Chapter 1 by the Editor is a useful introduction which provides an overview of the role of HPLC in biotechnology and a guide to subsequent chapters. Chapter 2, which deals with the design and synthesis of silica-based bonded phases for reversed-phase, ion-exchange and hydrophobic interaction chromatography, includes details for the purification of lysoxyme, ovalbumin and other proteins. Chapter 3 deals with nature of the sorption forces and the thermodynamics of protein adsorption. The next chapter addresses the important question of how much denaturation of proteins by organic solvents and consequent loss of biological activity occurs as a result of reversed-phase HPLC (RP-HPLC). The success of the technique depends very much on the particular protein-for example it can be used successfully for the isolation of human insulin without damage to the molecule, whereas with many other proteins reduced bioactivity may occur. This links in with Chapter 8 which deals in some detail with the use of analytical RF-HPLC in biosynthetic insulin production. Chapter 5 deals with the physical properties and applications of new synthetic hydrophilic vinyl polymer packing materials for industrial separation of biopolymers. Next follows a’ substantial chapter on the scale-up of protein chromatographic separation, which contains some introductory material on the basic principles of chromatography which would not have been out of place in the introductory chapter. The subject of Chapter 7 is the HPLC of antigenic proteins and vaccines. Chapter 9 deals very specifically with the application of RP-HPLC to the separation of human growth hormone, and a more general review of methods and mechanisms of high performance ion-exchange chromatography of proteins is the subject of the next chapter. In Chapter 11 the complementary nature of ion-exchange HPLC and reversed-phase HPLC for polypeptide purification is pointed out-complementary in the sense that their combined use can provide the optimal separation or purity assessment of a peptide. Hydrophobic interaction HPLC is the subject of Chapter 12, and Chapter 13 deals with high performance affinity chromatography (HPAC) or high performance liquid affinity chromatography (HPLAC) whichever one prefers to call it. The next chapter on the purification of synthetic oligonucleotides provides a change from other chapters, which seem to concentrate almost exclusively on peptides and proteins. At first glance, one might consider a single chapter on oligonucleotides to represent an imbalance in the book, but on closer inspection one quickly realizes that it is a very extensive and detailed chapter (108 pages, 248 references). The emphasis of most chapters on the application of HPLC to polypeptide purification reflects the enormous effort which goes into the production of polypeptides which are antigenically or physiologically active. The final chapter deals with the chemical synthesis of peptides and the contribution of mass spectrometry to their analysis. There is such a bewildering number of terms used in this book that a glossary of terms would not have been out of place and indeed it would have been welcomed. It does not help when alternative names for the same technique (gel filtration and size exclusion chromatography) are used in different chapters-the latter term is much preferred. All chapters of the book are well-referenced and are illustrated with clear line drawings and photographs. The book is very broad in coverage, both of techniques and applications. This is not a book for the newcomer to HPLC but I am certain that it will prove to be a valuable source-book to meet the needs of all workers in the field, whether they be analytical chemists, biochemists or chemical engineers. A. K. DAVIES Pharmaceutical Chemicals in Perspectbe: B. G. f59.00.
&mBEN
and H. A. WITTCOFF,Wiley, Chichester, 1989. Pages xviii + 518.
The aim of this book is to provide an overview of the pharmaceutical industry and its more important products. This is a somewhat ambitious goal to achieve within a mere five hundred pages. The book is divided into three parts. The first part includes as an introductory chapter a brief review of the chronological introduction of drugs, coupling this to statistical data on changing patterns of mortality. This is followed by a wide-ranging
1205
There are times also when I would have preferred a more detailed description of the underlying assumptions of a given technique, and more mathematical formulae, but these are relatively small matters. The book begins with revision of r- and F-tests, which leads naturally to simple analysis of variance techniques. Experimental design methods are then described for systems of increasing complexity. Factorial design then follows, and then fractional factorial design. The book concludes with a chapter on response surfaces. This is the right subject matter, well handled, and I recommend this book as a good introduction to chemometrics. C. J. GILMORE High Performance Liquid Chromatographyin Bioted~m-dogy:W. S. HANCOCK (editor), Wiley-Interscience, New York, 1990. Pages x + 564, E74.35. Lavoisier’s axiom that only pure substances would yield information of significance is as true today as it was in his day. We now possess the techniques to enable us to isolate well-defined compounds that satisfy the most stringent of purity. There can be no doubt that high performance liquid chromatography has proved to be one of the most important and rapidly advancing of these techniques. Since biotechnology is another branch of science which is making rapid strides and which offers considerable promise for the future, this book, which examines the various applications of HPLC to biotechnology is very timely indeed. It is worth noting that the majority of papers published recently which are concerned with HPLC have concentrated on biological applications. The book nicely reflects the multifaceted nature of the subject and the important interplay between its analytical and preparative aspects. The range of applications stretches from the nanogram scale, for characterization studies of novel proteins, to the kilogram amounts required for a manufacturing process. Chapter 1 by the Editor is a useful introduction which provides an overview of the role of HPLC in biotechnology and a guide to subsequent chapters. Chapter 2, which deals with the design and synthesis of silica-based bonded phases for reversed-phase, ion-exchange and hydrophobic interaction chromatography, includes details for the purification of lysoxyme, ovalbumin and other proteins. Chapter 3 deals with nature of the sorption forces and the thermodynamics of protein adsorption. The next chapter addresses the important question of how much denaturation of proteins by organic solvents and consequent loss of biological activity occurs as a result of reversed-phase HPLC (RP-HPLC). The success of the technique depends very much on the particular protein-for example it can be used successfully for the isolation of human insulin without damage to the molecule, whereas with many other proteins reduced bioactivity may occur. This links in with Chapter 8 which deals in some detail with the use of analytical RF-HPLC in biosynthetic insulin production. Chapter 5 deals with the physical properties and applications of new synthetic hydrophilic vinyl polymer packing materials for industrial separation of biopolymers. Next follows a’ substantial chapter on the scale-up of protein chromatographic separation, which contains some introductory material on the basic principles of chromatography which would not have been out of place in the introductory chapter. The subject of Chapter 7 is the HPLC of antigenic proteins and vaccines. Chapter 9 deals very specifically with the application of RP-HPLC to the separation of human growth hormone, and a more general review of methods and mechanisms of high performance ion-exchange chromatography of proteins is the subject of the next chapter. In Chapter 11 the complementary nature of ion-exchange HPLC and reversed-phase HPLC for polypeptide purification is pointed out-complementary in the sense that their combined use can provide the optimal separation or purity assessment of a peptide. Hydrophobic interaction HPLC is the subject of Chapter 12, and Chapter 13 deals with high performance affinity chromatography (HPAC) or high performance liquid affinity chromatography (HPLAC) whichever one prefers to call it. The next chapter on the purification of synthetic oligonucleotides provides a change from other chapters, which seem to concentrate almost exclusively on peptides and proteins. At first glance, one might consider a single chapter on oligonucleotides to represent an imbalance in the book, but on closer inspection one quickly realizes that it is a very extensive and detailed chapter (108 pages, 248 references). The emphasis of most chapters on the application of HPLC to polypeptide purification reflects the enormous effort which goes into the production of polypeptides which are antigenically or physiologically active. The final chapter deals with the chemical synthesis of peptides and the contribution of mass spectrometry to their analysis. There is such a bewildering number of terms used in this book that a glossary of terms would not have been out of place and indeed it would have been welcomed. It does not help when alternative names for the same technique (gel filtration and size exclusion chromatography) are used in different chapters-the latter term is much preferred. All chapters of the book are well-referenced and are illustrated with clear line drawings and photographs. The book is very broad in coverage, both of techniques and applications. This is not a book for the newcomer to HPLC but I am certain that it will prove to be a valuable source-book to meet the needs of all workers in the field, whether they be analytical chemists, biochemists or chemical engineers. A. K. DAVIES Pharmaceutical Chemicals in Perspectbe: B. G. f59.00.
&mBEN
and H. A. WITTCOFF,Wiley, Chichester, 1989. Pages xviii + 518.
The aim of this book is to provide an overview of the pharmaceutical industry and its more important products. This is a somewhat ambitious goal to achieve within a mere five hundred pages. The book is divided into three parts. The first part includes as an introductory chapter a brief review of the chronological introduction of drugs, coupling this to statistical data on changing patterns of mortality. This is followed by a wide-ranging