TIBTECH- FEBRUARY 1991 [Vol. 9] 69

O000//or BI]BBO/ Pure thinking about proteins

GUIDE TO PROTEIN PURIFICATION:

METHODS IN ENZYMOLOGY, VOL. 1 8 2

edited by Murray P. Deutscher, Academic Press, 1990. $49.95 (xii + 894 pages) ISBN 0 12 213585 7

I reviewed this guide with con- stantly changing impressions. After the initial shock of seeing the fam- iliar green livery replaced by a spiral bound paperback, there was a feeling of apprehension: the guide has more than 900 pages of text. This was tempered by awe at the names of 69 contributors, including many of 'the great and the good'. Somehow these had been induced to deliver copy by the evidently superhuman editor. Any idea that I was about to be bored was abolished by the freshness of Arthur Kornberg's introductory prose. This takes the mundane and often frankly tedious business of isolating proteins onto another plane: 'Don't waste clean thinking on dirty enzymes'. In the epilogue, other giants of biochemistry remind us of the inherited wealth {often taken for granted} that we possess through the study of purified pro- teins. Strong stuff for a practical guide.

But then this is more than just a guide. It is a complete work. The strategies described embody both classical and modern thinking. We are told what to do to set up a separation laboratory in classic terms (don't start with wall-to-wall FPLC). New biochemists sometimes neglect their roots in physical chemistry. The chapters on buffers, protein assay and quantitation will repay study. The editor has recog- nized that the everyday problems are protein desalting, concentration, recovery and storage. The advice here is comprehensive.

Excellent chapter follows chapter. Expression of proteins in pro- and eukaryotic systems; recovery of the same, from organelles, from mem- brane fractions, from inclusion bodies and even from the super- natant, all receive attention. The major separation methods are

described for all notable classes of protein (including HPLC, affinity methods, immunological methods, chromatofocusing, immobilized metal- ion chromatography and electro- elution).

Every reviewer tries to find some- thing that 'could have been done better'. Well, you wouldn't find much on expression of proteins in that useful commercial organism, yeast - but you probably wouldn't

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High standards PROTEIN PURIFICATION-- PRINCIPLES,

HIGH RESOLUTION METHODS AND

APPLICATIONS

edited by Jan-Christer Janson and Lars Ryden, VCH VerlagsgeseUschaft, 1990. £45.00 (xiii + 243 pages) ISBN 1 85166 401 7

This work represents a compilation of 'current' high-resolution methods for protein purification, co-written by researchers in the field and Pharmacia LKB Ltd. Each chapter has been reworked by the editors to ensure a pleasantly uniform presen- tation. Emphasis has been placed on techniques, and the techniques de- scribed cover the theory and the practical considerations concluding, in most cases, with a series of re|- evant applications. A balanced mix- ture of specific and more general references is given at the end of each chapter.

The book is divided into three parts. Part I is a short introductory overview on how to purify a protein from starting material, through frac- tionation procedures, to the final product. Useful suggestions are given on protein monitoring and handling, and a comprehensive list of essential laboratory equipment is included. This part will be of value to the newcomer to this subject.

The purification methods which fill most of the book cover chroma-

buy this book for that. The section on protein characterization is incom- plete, but it's a bonus in this work anyway and the chapter on amino acid analysis is excellent.

In summary, this is possibly the most important, comprehensive and affordable work on protein purifi- cation to have appeared in recent years. Librarians who want to retain it as a reference work will have to put it behind glass and turn a page each day. This reviewer is going to keep his copy at home.

MICHAEL i. GEISOW

Biodigm, 115 Main Street, East Bridgeford, Nottingham NG13 8NH, UK.

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for resolution tography (Part II) and electrophoresis (Part HI). Key concepts on stationary- phase techniques and general chromatographic theory ease the reader into Part II with an intro- ductory chapter. Following chapters cover gel filtration, isoelectric focus- ing (IEF), chromatofocusing, re- versed-phase-HPLC, hydrophobic interaction chromatography (HIC), covalent and affinity chromato- graphy concluding with affinity partitioning and the benefits of two- phase systems and counter-current distribution. The experienced re- searcher may cover old ground here but each chapter is well supported by clear diagrams and illustrations, as well as comprehensive tables on gel media, buffers, etc. Much time is spent discussing 'standard' chroma- tography techniques with little dis- cussion on micro techniques. High resolution is often linked nowadays with small amounts of protein. Pharmacia LKB are addressing this problem with the introduction of their 'SMART' system and this will no doubt be considered for a chapter in a later edition.

Part III continues in a similar vein covering SDS--PAGE, IEF and im- munoelectrophoresis. Conventional methodologies are covered but it is strange that a book of this type does not include more discussion on prep- arative electrophoretic techniques.

70 T~BTECH- FEBRUARY 1991 rVol. 9]

[For example the Biorad Rotofor, Rainin Protein Fractionator and Applied Biosystems Inc. (ABI) 230 HPEC {High Performance Electro- phoresis Chromatography) system could all be considered here. Per- haps this is a case of technology moving faster than the published word?] Protein mapping of complex mixtures is considered in a chapter on two-dimensional PAGE with a complementary examination on pro- tein recovery and blotting tech- niques. These chapters are all well cross-referenced. The book concludes

with a chapter on capillary electro- phoresis and the many methods associated with this technique. It emphasizes the benefits in terms of speed and high resolution but gives few examples of specific appli- cations. The authors only touch on the usefulness of this technique which no doubt will become as popular as HPLC and PAGE.

There is an obvious need for high- resolution methods for purifying proteins. This is a rapidly expanding field and the authors must be con- gratulated on producing a well-

balanced collection of popular tech- niques, highlighting both theoretical and practical considerations. The book offers a sound foundation for the newcomer, and an excellent reference for any experienced researcher in the field.

ROY HARRIS

Protein ChemisL~ Section, Delta Biotechnology Ltd, Castle Court, Castle Boulevard, Nottingham NG7 1FD, UK.

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A laboratory guide for novice and expert BIOTECHNOLOGY: A LABORATORY

COURSE

by J. M. Becker, G. A. Caldwell and E. A. Lachgo, Academic Press, 1990. $29.95 (xviii + 232 pages) ISBN 0 12 084560 1

If you have just been appointed as a lecturer, or are looking to update and improve laboratory courses on microbiology and biotechnology, then this is the book for you. You do not even have to provide practical schedules, just recommend to students that they buy this book! Although it is designed as a labora- tory course, the book is also of use to new workers in the fields of mol- ecular biology and microbiology.

The manual is based on an intro- ductory laboratory course for senior undergraduates and first-year gradu- ate students in biological sciences at The University of Tennessee. It uses a series of experiments with Escheri. chia coil and Saccharomyces cere- visiae to teach everything from the basic principles of pH measurement, handling micropipettes and aseptic technique to the more advanced methods of plasmid isolation, electrophoresis and the use of DNA probes. These are just the types of experiments and the methods that students need to be taught as an introduction to molecular biology.

A short introduction to each exer- cise/practical section includes an overview of the methods and some of the theory behind the procedures. A brief list of references at the end directs the student to further reading on the subject. A list of specialist

reagents and equipment is also given, useful for a course organizer checking the practicability of using the exercise. The procedures them- selves are well written and easy to follow, designed for the novice, with clear, well-annotated diagrams where necessary. Questions at the end of each section help to consolidate the skills and information provided in the practical session.

As a laboratory-based course, this manual is very useful to students and course organizers, but it is the inclusion of extra sections and appendices which gives the book a more general appeal. There is a great emphasis on safety, in particular the handling and disposal of toxic chemicals and microorganisms, with additional notes on general labora- tory safety. There are sections on

record-keeping and laboratory note books. The appendices include in- formation on buffer preparation, sterilization, handling of restriction enzymes, to name but a few, thus making the book a good general reference book for anyone unused to handling microbes. For the new students, there is a glossary which should help them to understand the words and jargon that experienced laboratory workers routinely use, unaware that the uninitiated may not understand. An index is in- cluded, and the book is ring-bound so it will open fiat on the bench. It is a well-planned book and would be extremely useful to both the novice and the expert.

KATE GRAEME-COOK

Department of Biology, Birkbeck College, Malet Street, London WCIE 7I-IX, UK.

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