Methods for Affinity-Based Separations of Enzymes and Proteins

225 Pages·2002·2.18 MB·English

by  Munishwar N. Gupta

Methods and Tools in Biosciences and Medicine

Microinjection,
edited by Juan Carlos Lacal, Rosario Perona and James Feramisco, 1999
DNAProfilingand DNAFingerprinting,
edited by Jorg Epplen and Thomas Lubjuhn, 1999
Animal Toxins- Facts and Protocols,
edited by Herve Rochat and Marie-France Martin-Eauclaire, 2000
Methods in Non-Aqueous Enzymology,
edited by Munishwar Nath Gupta, 2000
Techniques in Molecular Systematics and Evolution,
edited by Rob DeSalle, Gonzalo Giribet andWard Wheeler, 2002
Methods for Affmity-based Separations ofEnzymes and Proteins,
edited by Munishwar Nath Gupta, 2002
Methods for Mfinity-Based
Separations of Enzymes and
Proteins
Edited by
Munishwar Nath Gupta
Springer Basel AG
Prof. Dr. Munishwar Nath Gupta
Department of Chemistry
Indian Institute of Technology, Delhi
Hauz Khas, New Delhi 110016
India
Library of Congress Cataloging-in-Publication Data
Methods for affinity-based separations of enzymes and proteins / edited by Munishwar Nath
Gupta.
p.cm. - (Methods and tools in biosciences and medicine)
Includes bibliographical references and index.
ISBN 978-3-7643-6305-5 ISBN 978-3-0348-8127-2 (eBook)
DOI 10.1007/978-3-0348-8127-2
1. Affinity chromatography-Laboratory manuals. 2. Proteins-Separation-Laboratory
manuals. 3. Enzymes-Separation-Laboratory manuals. I. Gupta, Munishvar Nath. 11
Series.
Deutsche Bibliothek Cataloging-in-Publication Data
Methods for affinity-based separations of enzymes and proteins / ed. by Munishwar Nath
Gupta. - Springer Basel AG, 2002
(Methods and tools in biosciences and medicine)
ISBN 978-3-7643-6305-5
The publisher and editor can give no guarantee for the information on drug dosage and
administration contained in this publication. The respective user must check its accuracy by
consulting other sources of reference in each individual case.
The use ofregistered names, trademarks etc. in this publication, even ifnot identified as such,
does not imply that they are exempt from the relevant protective laws and regulations or free
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This work is subject to copyright. All rights are reserved, whether the whole or part of the
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@ 2002 Springer Basel AG
Originally published by Birkhäuser Verlag in 2002
Softcover reprint ofthe hardcover Ist edition 2002
Printed on acid-free paper produced from chlorine-free pulp. TCF
00
Cover illustration: An affinity based separation of a protein (with the friendly permission of
Prof. Gupta)
ISBN 978-3-7643-6305-5
987654321 www.birkhasuer-science.com
Contents
List ofContributors...................................................................................... VII

Preface IX

1 Aftlnity-Based Separation: An Overview........................................... 1

Munishwar N. Gupta and Ipsita Roy

2 Atrmity Chromatography..................................................................... 16

Nikolaos E. Labrou

3 Expanded BedAtrmity Chromatography........................................... 29

Arvind Lali

4 P erfusionAtrmity Chromatography for Rapid Antibody

Separations........................................................................................... 65

Robert L. Fahrner and Gregory S. Blank

5 A trmity-Based Interactions on Disks for FastAnalysis,

Isolation and Conversion ofBiomolecules... 82

Laure G. Berruex and Ruth Freitag

6 Sugars as Atrmity ligands 115

Posettihalli R. Satish and Avadhesha Surolia

7 Macroaffinity ligands in Bioseparation 130

Ipsita Roy and Munishwar N. Gupta

8 Cellulose-Binding Fusion Proteins...................................................... 148

Alisdair B. Boraston, BradleyW. McLean, Mojgan Kavoosi,

CharlesA. Haynes and Douglas G. Kilburn

9 Two-PhaseAtrmity PartitioningofAnimal Cells: Implications

ofMultipoint Interactions 163

Ashok Kumar, Masamichi Kamihira and Bo Mattiasson

10 Atrmity-Based Reverse Micellar Separations.................................... 181

Motonari Adachi

11 Biosensors: An Emerging Toolin Bioanalytical Research­

AKinetic Analysis ofthe Analyte-Receptor Interactions in

Biosensors............................................................................................. 195

Anand Ramakrishnan and Ajit Sadana

Guide to Solutions 217

Guide to Enzymes 217

Guide to Protocols 217

TroubleshootingGuide 218

Index.............................................................................................................. 221
VII
List of Contributors
ADACHI, MOTONARI, Institute ofAdvanced Energy, Kyoto University, Uji, Kyoto
611-QOll,Japan; email: [email protected]
BERRUEX, LAURE G., Centre of Biotechnology, Faculty of Basic Sciences, Swiss
Federal Institute ofTechnology, CH-1015 Lausanne, Switzerland
BLANK, GREGORY S., Department ofRecovery Sciences, Genentech, Inc., 1 DNA
Way, South San Francisco, CA 94080, USA
BORASTON, ALISDAIR 8., The Biotechnology Laboratory, University of British
Columbia,VancouverBC, CanadaV6T1Z3
FAHRNER, ROBERTL., DepartmentofRecoverySciences, Genentech, Inc., 1DNA
Way, SouthSan Francisco, CA94080,USA; email: [email protected]
FREITAG, RUTH, Centre of Biotechnology, Faculty of Basic Sciences, Swiss
Federal Institute of Technology, CH-1015 Lausanne, Switzerland; email:
[email protected]
GUPTA, MUNISHWAR N., CheInistry Department, Indian Institute ofTechnology,
Delhi, Hauz Khas, New Delhi 110016, India; email: [email protected]
HAYNES, CHARLES A., The Biotechnology Laboratory, University of British Co­
lumbia,Vancouver BC, CanadaV6T 1Z3
KAMIHIRA, MASAMICHI, Department of Biotechnology, Graduate School of
Engineering, Nagoya University, Chikusa-ku, Nagoya 464-8601, Japan
KAVOOSI, MOJGAN, Department of CheInical and Biological Engineering, 2216

Main Mall, University ofBritish Columbia, Vancouver, CanadaV6T 1Z4

KILBURN,DOUGLASG.,TheBiotechnologyLaboratory,UniversityofBritishColum­

bia,VancouverBC,CanadaV6T1Z3;email: [email protected]
KUMAR, ASHOK, Department of Biotechnology, Center for CheInistry and Che­
Inical Engineering, Lund University, S-22100 Lund, Sweden
LABROU, NIKOLAOS E., Laboratory ofEnzyme Technology, Department ofAgri­
culturalBiotechnology,AgriculturalUniversityofAthens,GR-11855-Athens,
Greece; email: [email protected]
LALI, ARVIND, CheInical Engineering Division, Department of CheInical Tech­
nology, University of Mumbai, Matunga, Mumbai 400 019, India; email:
[email protected]
w.,
McLEAN, BRADLEY The Protein Engineering Network of Centres of Excel­
lence,PENCEInc., NationalBusinessCentre,750HeritageMedicalResearch
Centre, Edmonton, Alberta, Canada T6G 2S2
MATTIASSON, Bo, Department of Biotechnology, Center for CheInistry and
CheInical Engineering, Lund University, S-22100 Lund, Sweden; email:
[email protected]
RAMAKRISHNAN, Anand, CheInical Engineering Department, University of
Mississippi, University, MS 38677-1848, USA; email: [email protected]
RoY, IpSITA, CheInistryDepartment, Indian Institute ofTechnology, Delhi, Hauz
Khas, New Delhi 110016, India
VIII
SADANA, AnT, Chemical Engineering Department, University of Mississippi,
University, MS 38677-1848, USA; email: [email protected]
SATISH, POSETTIHALLI R., Molecular BiophysicsUnit, Indian Institute ofScience,
Bangalore 560012, India
SUROLIA, AVADHEsHA, Molecular Biophysics Unit, Indian Institute of Science,
Bangalore 560 012, India; email: [email protected]
IX
Preface

Shapeyourhearttofrontthehour,
butdream notthatthehourwill lost.
Alfred Tennyson
Locksley Hall Sixty YeO!, After
Numerous advances in the life sciences have necessitated the need to develop
gentle, quick and economical methods for separation of proteins/enzymes.
Affmity-based separations, with their high selectivity, have thus come to be
used more frequently. In the beginning, we just had atrmity chromatography.
Today we have several strategies that employ atrmity interactions for separa­
tion and analysis. This book is an attempt to provide a short introduction and
illustrativeprotocolsfor someimportantandcurrentaffinity-basedapproaches
intheareaofdownstreamprocessingofenzymes/proteins.Today,peoplefrom a
variety of disciplines have to purify proteins. It is hoped that this book will
provide some quick insight into the various available options and guide even a
novice (in the area ofprotein separation) to obtainhis/her target protein at the
level ofpuritythat he/she desires.
Anoverview(ChapterI)ofsuchmethodsisfollowedbythedescriptionofvarious
techniques in which affinity interactions have been built-in. This includes ex­
panded bed chromatography (Chapter Ill), perfusion chromatography (Chapter
IV),monoliths(ChapterV),two-phaseextraction(ChapterIX)andreversemicellar
extraction (ChapterX). Some novel adsorbents are describedin Chapters II and
VII.Thelatterfocusesonmaterialswhichhaveaninherentaffinitytowardssome
useful enzymes and can also be used for separation in free solutions. Sugar­
bindingproteins are animportantclassofproteinsandChapterVIdescribesthe
preparation ofaffinityadsorbentsfor these proteins. ChapterVIIIillustratesthe
frequentlyusedapproachof'tags'tofacilitate proteinrecovery. Thelastchapter
(Chapter XI) is a little different, inasmuch as it has no experimental protocol. It
dealswithreal-time analysisofaffinityinteractionsusingfractals.
While the methods describedin all the other chaptersrelate to separation of
proteins, Chapter IX focuses on separation of animal cells. However, as the
authorsstate,the approachoutlinedisequallyvalidfor proteinseparation. It is
justthattheauthorshavechosentoillustratethetechniquewithamorecurrent
and exciting example.
Iamgrateful to the various experts who have agreed to share their valuable
experience through these chapters. Much of my interest in bioseparation
originated in the enjoyable collaboration with Prof. Bo Mattiasson (Sweden).
Those few beautiful summers spent in his laboratory at Lund defmitely con­
tributed to increasing my affinity for this area.
Iwould also like to acknowledge:
• People at Birkhauser Verlag, notably Ms. Katrin Serries, Dr. Hans Detlef
Kliiberand Ms. Daniela Brunnerfor their patience and painstakingefforts.
x
• Department of Science and Technology, Department of Biotechnology,
Council of Scientific and Industrial Research (Extramural Division and
Technology Mission on Oilseeds, Pulses and Maize) (all Govt. of India
organizations) for funding my research work over the years.
• lIT Delhi for providing a modest book-writing grant.
• Myco-workersin myresearch group: Dr. S. K. Khare, Dr. A. Sharma, Dr. S.
Teotia, Dr. I. Roy, Ms.S. Sharma and Ms.S. Jain. One ofthem, Dr. Roy, has
also co-authoredtwo ofthe chapterswithme. Apartfrom that,shehas also
been a great help at allstages- from inceptionto the final editing- during
the preparation ofthe book.
• My wife, Sulbha, and my son Chetan, who "managed without me" quite
often during the period ofmy involvementwith this book.
Iwould welcome any comments from the readers/users.
May, 2001 M.N.Gupta