Antibody Expression and Engineering

162 Pages·1995·15.99 MB·English

by  Henry Y. Wang and Tadayuki Imanaka (Eds.)

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In Antibody Expression and Engineering; Wang, H., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
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In Antibody Expression and Engineering; Wang, H., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
A CS S Y M P O S I UM SERIES 604
Antibody Expression
and Engineering
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95- Henry Y. Wang, EDITOR
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pubs.acs.org ust 18, 1995 | d Osaka University
12 | http://Date: Aug Developed from a symposium sponsored
July 17, 20 Publication by thoef atDht ietvh iAseim o2n0er7 oitcfha Bn Ni oCacthhioeenmmaiilcc aMall eSTeoetcicnihegtny o,l ogy
San Diego, California,
March 13-17, 1994
American Chemical Society, Washington, DC 1995
In Antibody Expression and Engineering; Wang, H., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
Library of Congress Cataloging-in-Publication Data
Antibody expression and engineering / Henry Y. Wang, editor, Tadayuki
Imanaka, editor.
p. cm.—(ACS symposium series, ISSN 0097-6156; 604)
"Developed from a symposium sponsored by the Division of
Biochemical Technology at the 207th National Meeting of the American
Chemical Society, San Diego, California, March 13-17, 1994."
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0 Includes bibliographical references and indexes.
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4.f ISBN 0-8412-3314-4
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5-0 1. Monoclonal antibodies—Biotechnology—Congresses.
9 2. Immunogenetics—Congresses. 3.I mmunoglobulin genes—
9
1 Congresses. 4. Hybridomas—Congresses. 5. Gene expression—
bk- Congresses.
1/
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0 I. Wang, Henry Y., 1951- . II. Imanaka, Tadayuki, 1945- .
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0. III. American Chemical Society. Division of Biochemical Technology.
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doi: ICVa.l ifA.)m Ver.i cSaenr ieCsh. emical Society. Meeting (207th: 1994: San Diego,
pubs.acs.org ust 18, 1995 | 4ET.Px [p2GDr4eeN8sn.sL6ei5oMti.nMc:— E615cn.oA gAni5ngn6ertee ibsris1one9dgs9i—.e 5s3— m. egGtheenonedtesi—ctisc—c Eocxnopgnrreegssrsseiesossne. —s.Q cW2o.n gG5r7ee5sn seAes 6. 29 1995]
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July 17, 20Publication TChoipsy briogohkt ©is p19ri9n5te d on acid-free, recycled paper.

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In Antibody Expression and Engineering; Wang, H., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
1995 Advisory Board
ACS Symposium Series
Robert J. Alaimo Cynthia A. Maryanoff
Procter & Gamble Pharmaceuticals R. W. Johnson Pharmaceutical
Research Institute
Mark Arnold
University of Iowa Roger A. Minear
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1D University of Wisconsin at Madison Concurrent Technologies Corporation
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Madeleine M. Joullie Michael D. Taylor
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Lawrence P. Klemann William C. Walker
Nabisco Foods Group DuPont
Douglas R. Lloyd Peter Willett
The University of Texas at Austin University of Sheffield (England)
In Antibody Expression and Engineering; Wang, H., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
Foreword
THE ACS SYMPOSIUM SERIES was first published in 1974 to
provide a mechanism for publishing symposia quickly in book
form. The purpose of this series is to publish comprehensive
1 books developed from symposia, which are usually "snapshots
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w in time" of the current research being done on a topic, plus
04.f some review material on the topic. For this reason, it is neces
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5-0 sary that the papers be published as quickly as possible.
99 Before a symposium-based book is put under contract, the
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0.1 papers are excluded at this point, and others are added to
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d paper is peer-reviewed prior to final acceptance or rejection.
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In Antibody Expression and Engineering; Wang, H., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
Preface
ANTIBODIES ARE MULTTDOMAIN GLYCOPROTEIN molecules of intense
interest to the biotechnology community. Antibodies are immunoglobulin
molecules of the immune system developed by higher organisms to com
bat the invasion of foreign substances (antigens). Each antibodty has
specific activity for the foreign material that elicits its synthesis. In
nature, each antibody is made by a different line of lymphocytes and their
1 derived plasma cells in a specific animal. If one could pick out such a cell
0
pr0 making a single specific antibody and grow it in a culture, then the cell's
04. progeny, or clone, would be a source of large amounts of identical anti
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5-0 body against a single antigenic determinant: a monoclonal antibody
99 (MAb*). Unfortunately, antibody-secreting cells cannot be maintained
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k- well in a culture medium.
b
21/ In 1975, Kohler and Milstein fused mouse myeloma cells with lym
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0.1 phocytes from the spleen of mice immunized with a particular antigen.
1
oi: The resulting hybrid myeloma, or "hybridoma," cells expressed both the
g 5 | d lymphocytes' property of specific-antibody production and the immortal
s.or199 character of myeloma cells. Since then the hybridoma technology has
s.ac18, been widely adopted as a method of choice for the preparation of MAbs
ubst to a wide spectrum of antigens. Various hybridomas can be cloned and
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Some of the real or potential applications of monoclonal antibodies
will require kilogram quantities of highly purified antibody. Therefore,
cost-efficient methods for producing large quantities of highly purified
monoclonal antibodies must be developed. Process development will be
an important factor for the successful commercialization of any of the
therapeutic monoclonal antibodies. Through better understanding of the
nutritional requirements of these cells, many improvements in culture
media and feeding strategies have been developed. Costly and undefined
serum components have been eliminated from media formulation without
*A number of abbreviations for monoclonal antibodies are used in this book: MAb, Mab, Abs,
MoAbs. They all denote the same thing.
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In Antibody Expression and Engineering; Wang, H., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
impairing the immunoglobulin secretion or the consistency of the anti­
body properties. Development of serum-free media has also made the
antibody-manufacturing process more economically competitive.
Many antibodies have the potential to be the most economically signi­
ficant products derived from mammalian cells. They are already being
used extensively in diagnostic assays and their therapeutic potential is
increasing with the development of affinity purification systems and in
vivo imaging. The immunotherapeutic application uses monoclonal anti­
bodies either as so-called "magic bullets" to destroy malignant tissue
(cancer therapy) or as target-specific drug delivery agents (for immuno­
logical diseases).
Recent progress in our ability to genetically manipulate and alter the
antibody molecules and then express the whole antibody molecule or its
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1/b In the first few chapters of this book, recent advances in antibody expres­
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10. dies based on traditional myeloma cells are described. Using a case-study
oi: approach, David Robinson and his colleagues show how gene amplifica­
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g 5 | tion, medium optimization, and process development can be used to
s.or199 optimize a humanized anti-CD18 antibody. Almost 1 g/L of monoclonal
s.ac18, antibody can be routinely produced in an optimized fed-batch culture.
pubust Sawada and Kitano describe the development and characterization of a
p://Aug human monoclonal antibody against hepatitis Β virus surface antigen
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Chinese hamster ovary (CHO) cells. Almost 800 mg/L of antibody was
achieved in a 100-L fermentor in 6 days. This is quite competitive with
the traditional myeloma hosts.
Genetic shuffling of antibody domains and other active molecules can
also be used to create new chimeric molecules with various combinations
of binding and effector functions and novel antigen-binding molecules.
Similarly, recombinant DNA techniques have now been used to produce
different antibody fragments with reduced molecular mass, such as Fab,
Fv, or single-chain antigen-binding molecules. These engineered mono­
clonal antibodies and their corresponding fragments have the potential to
expand on various clinical applications, such as diagnostic imaging and
immunotherapeutics, that require smaller molecules for deeper penetra­
tion or with altered binding properties. The next few chapters are devoted
χ
In Antibody Expression and Engineering; Wang, H., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
to the expression of monoclonal antibody and its fragments in other non-
mammalian host systems. Potter et al. demonstrate the use of baculovirus
expression system to produce various forms of hybrid or chimeric antibo
dies of IgG, IgM, and IgA isotypes in insect cells. Julian Ma reports on
the progress of expressing antibody fragments in cultivated plant cells and
plants.
Microbial gene expression systems have also been used routinely to
produce antibody fragments (such as Fab and Fv) consisting of one or
two antibody chains. Co-expressing antibody light and heavy chains in
various microbial hosts have been demonstrated since 1984. David Fil-
pula and his colleagues review the potential of using the Escherichia coli
expression system to overproduce a single-chain Fv protein with human
IgM Qx specificity. Better and Nolan describe the construction of a fam
ily of immunofusion proteins linking various antibody fragments of the
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4. called gelonin in E. coli. Some of these fusion proteins are as potent as
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5-
9 toxins. S. L. Wong and his group have shown that a multiple-protease-
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1 ment. Nyyssonen and Keranen discuss the use of cellulase-producing Tri-
0.
doi: 1 cthheo dceerlmluala rseee gseeni ea. s a promising host to produce a Fab fragment fused to
ubs.acs.org st 18, 1995 | psyronmtIhnie ssitinhsg.e Frfieoemlrd a ai onmfi nocgra etc ahslypapteitccei rfasicn, taiLbpaopvdleiiyce asa tinaodnn d,J Tatnh. deImair aranepavpkiealiw'cs a tgthiroeon usep x icnhi atoivnregg asaunncidc
p://pAugu cessfully generated a light chain of a MAb against porphyrin that still
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July 17, 20 Publication bbamneion dlaa en ecctnu ohglmeoi nsphetr aeesvrhyiense tngebs meiuvcssoe ia mnhngead n tkarda nbfdoeoiwwotik nop. n otahtIlern nochutryiegabahlrs iaidnXpogp-mlrlyaiac,y a faticinnrodeyn ssdtn.ae oltIlnaotit glrissar adnopifoth ittyo hi nenaa tnlea dnnr NdeticebModom dRtoy
spectroscopy. Thus, quantitative relationships between structure and
function can now be estimated and manipulated through genetic engineer
ing. Similarly, screening for novel antibodies through the generation of
combinatorial libraries using filamentous phage and other microbial anti
body expression systems is becoming routine. Therefore, the advances in
genetic manipulation and process development discussed in this book
coupled with other advances will propel the field of antibody engineering
to a new horizon.
Acknowledgments
We gratefully acknowledge the assistance of many anonymous reviewers
xi
In Antibody Expression and Engineering; Wang, H., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
who helped us carry out in-depth reviews and supplied important and
constructive criticism of the original manuscripts. We also acknowledge
the patience and support from the staff of the ACS Books Department,
particularly Barbara Pralle, that made this book a reality.
HENRY Y. WANG
Department of Chemical Engineering
University of Michigan
Ann Arbor, MI 48109-2136
TADAYUKI IMANAKA
Department of Biotechnology
Faculty of Engineering
Osaka University
1
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4. Osaka 565, Japan
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In Antibody Expression and Engineering; Wang, H., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1995.