Bioseparations: Downstream Processing for Biotechnology [1 ed.] 0471847372, 9780471847373

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01911-280780

BIOSEPARATIONS Downstream Processing for Biotechnology Paul A. Belter E. L Custer Wei-Shou Hu

A Wiley-lnterscieni JOHN WILEY & SONS

New York

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Chichester

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Brisbane

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Toronto

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Singapore

Marchetti, David Schisla, Steve Trank, and Jim Yang. Our colleagues at the Upjohn Company gave years of stimulus and criticism, each as needed. H. Ted Davis and Michael Flickenger of the University of Minnesota provided administrative and financial support. Finally, Julie Prince, who typed the manuscript, has truly lived up to her surname. Paul A. Belter E. L. Cussler

V/ei Shou Hu

To the Faculty of the University of Minnesota.

A Wiley-Interscience

January. 1988

Publication

Copyright © 1988 by John Wiley & Sons, Inc. All rights reserved. Published simultaneously in Canada.

Reproduction or translation of any part of this work \ beyond that permitted by Section 107 or 108 of the 1976 United States Copyright Act without the permission of the copyright owner is unlawful. Requests for permission or further information shquldjbc addressed to the Permissions Department, John Wiley & Sons, I nV - -ft Library of Congress Cataloging in Publication Data:

Belter. Paul A.. 1920Biosepa rations: downstream processing for biotechnology/ Paul A Belter. E.L. Cussler, Wei-Shou Hu. p.

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"A Wiley- In lerscience publication.” Includes index. ISBN 0-471-84737-2: $35.00 (est.) 1« Biomolecules— Separation. 2. Biotechnology

I. Cussler. EL. II. Hu. Wei-Shou 1951- III. TP248.25.S47B45 1988 660 '.63-de19 Printed and bound

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Technique. *

i 87-20292 CIP

the United States of America

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II

Preface

;eparatioi purification of This short book biochemicals. Large amounts of these biochemicals can be mai rryxihe land KVmethods of genetic engineering, including cloned microo %jed anti bridomas. After these biochemicals are made, thpy than the purified. These separations are difficult and f initial manufacture of the biochemicals. e parallel cultures This book has almost no prede life science hinges on of life science and engineering biochemistry, microbjQlo and jAhat of engineering includes mass cultures are reflected in two almost transfer and unit opeYah completely iscijflines, where students in one discipline have trouble taki in thy other. This is especially true of chemical ses in chemical separations routinely imply four or engin n8’ mor [al prerequisites. v\triW to write this book as a bridge between these two cultures, a^imple introduction to bioseparations for scientists with no back und in engineering and for engineers with no background in biology. As such, it includes substantial simplification. It may occasionally v

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Preface

appear biologically naive and mathematically trivial, but such appearances are part of most introductions. We have assumed a rudimentary knowledge of integral calculus in writing the book. While we recognize that some trained in the life sciences may not have this knowledge, we make no apology for this. In fundamental biotechnology, we want to know if we can make a desired product. Answering that question- often requires no mathematics. In commercial biotechnology, we already know that we can make the product; we want to know how much we can make and at what cost. Answering this commercial question usually implies mathematics. The book can be used by any scientifically trained individual interested in this field. However, we have used drafts of this book as the basis for a successful course taken by seniors and first year graduate students. The seniors are largely chemical engineers with few courses in biology. The graduate students have included those trained in chemistry, microbiology, agriculture, food science, and civil engineering. Other students who have returned from industrial careers have almost forgotten much' of their original training. All these students have worked hard, and their efforts to learn have spurred our revisions of this material. Our course treats the material in the order shown in the table of contents. We give 45 lectures, so the course fits into one academic semester. Wc begin the semester with two or three lectures reflecting specific uncer¬ tainties in the students’ preparation. For our chemical engineering students, we review biological materials, using the outline in Appendix A. Wc then give two lectures on existing overall processes, asking the students to divide these into the four most common steps: removal of insolubles, product isolation, purification, and polishing. With these four steps as a template, students design belter processes, even when they depart from the template. We use these four steps to organize the remaining material. We spend about five lectures on each of the following: filtration, centrifugation, extraction, adsorption, elution chromatography, and crystallization. Wc lecture less on cell, disruption, precipitation, ultrafiltration, and drying, partly because of the pressure of time and partly because these processes arc dominated by experiment rather than analysis. We rarely do justice to ancillary operations. Our students work hard and enjoy the course. Many helped us in writing the book. Michael Ladisch of Purdue Univer¬ sity and Joseph A. Shaeiwitz of West Virginia University read the manuscript, caught our more blatant errors, and made many practical suggestions. Students who were a great help include Daniel Lavin, Marcelo I

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Contents

Notatioi x

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An Overview of Bioseparatio 1.1 Characteristics of IOS 1.2 An Idealized Proc 1.3 Conclusions,

LUBLES

PART Iz REM

ion 2/ Filtrati t for Conventional Filtration, 14 2.1 Equip 17 2.2 Preke Theory for Filtration, 22 2.3 linuous Rotary Filters, 30 2.4 ratory Tests, 35 icJofiltration, 39

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Conclusions. 41