Biological Sciences
Monoclonal Antibodies
Monoclonal antibodies are laboratory-produced molecules designed to mimic the immune system's ability to fight off harmful pathogens. They are created by cloning a unique immune cell to produce large quantities of a single type of antibody. Monoclonal antibodies have various applications in medicine, including targeted cancer therapies, autoimmune disease treatments, and diagnostic tests.
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11 Key excerpts on "Monoclonal Antibodies"
eBook - PDF- Alberto Macario(Author)
- 2012(Publication Date)
- Academic Press(Publisher)
The manufacture of large quantities of antibacterial monoclonal anti-bodies for routine and widespread use in many areas (medicine, dentistry, veteri-nary sciences, industry, and biotechnology) is evolving into an industry of its own. The tactics for industrial production with commercial purposes are different from those in basic research. Also research and development endeavors in the Introduction xxiii industrial world differ from research efforts in the academic environment (see Volume II). D. Genetic Engineering Monoclonal Antibodies are instrumental in genetic engineering, first to identify useful antigens, for example, those inducing protective immunity, and then to help in the cloning of the genetic codes for these antigens in the preparation of vaccines or diagnostic kits (37,54). E. Biochemistry and Molecular Studies Elucidation of the chemical composition and structure of the antigen molecule bearing the determinant recognized by a monoclonal antibody, and eventually of the determinant itself, should become an important part of chemoimmunotax-onomy and other molecular studies (4,6,10,16,18,23,44,53,54,58,61,65) (see Chapters 1, 6-8, and Volumes II and III). For this purpose, panels of mono-clonal antibodies show extraordinary resolution power, especially if the fine (molecular) specificity of the antibodies is known. In this case, the antibodies constitute a set of high-precision tools for probing molecular markers and for detecting these markers in a variety of materials. One can foresee the occurrence of Monoclonal Antibodies specific for a marker of a strain, or species, or higher taxon. One can also envisage the use of Monoclonal Antibodies of predefined mo-lecular specificity for tracing molecular signatures left by a given strain in other microorganisms, subcellular structures (e.g., mitochondria), and materials from ecologic niches, such as fossils and sediments, and from culture superna-tants (see Chapter 11).
- Fazal Ahmad(Author)
- 2012(Publication Date)
- Academic Press(Publisher)
Monoclonal Antibodies—Production and Uses
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Monoclonal Antibodies: THE PRODUCTION OF TAILOR-MADE SEROLOGICAL REAGENTS
Dale E. Yelton, Pallaiah Thammana, Catherine Desaymard, Susan B. Roberts, Sau-Ping Kwan, Angela Giusti, Donald J. Zack, Roberta R. Pollock and Matthew D. Scharff, Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York U.S.A.Publisher Summary
Immunoassays have been used to detect, quantitate, and localize small amounts of macromolecules in complex biological mixtures. Monoclonal Antibodies are chemically defined reagents. Monoclonal Antibodies have many benefits over conventional antisera. In addition to providing homogenous reagents that can be generated in large amounts and replenished whenever needed, particular monoclonals can be selected. Once a hybridoma producing a desired antibody has been identified, hundreds of milligrams of that antibody can be generated. These benefits have stimulated investigators, in all areas of biology, to generate Monoclonal Antibodies against the particular antigen that they are studying. Monoclonal Antibodies can also be tailor–made by identifying subclones containing deletions in one or another domain. Such deletion variants have been generated from an IgG2b hybridoma producing antibody that reacts with hapten p-azophenylarsonate. Useful monoclonals can be made even more effective by isolating subclones that have undergone mutations or rearrangements.I. INTRODUCTION
Immunoassays have been used successfully for years to detect, quantitate, and localize small amounts of macromolecules in complex biological mixtures. While such assays can be made both sensitive and specific, certain properties of conventional antiserums have limited their usefulness, especially for routine diagnosis and therapy. Perhaps the most important of these limitations has been the limited supply of useful antiserum against weak immunogens. This is in part due to the heterogeneity of the immune response which results in each antiserum being a mixture of antibodies with varying affinity, cross reactivities, and effector functions. The particular mix of antibodies or predominance of a subset of antibodies produced by an animal at a certain time in its immune response may be useful for a specified purpose. However, as that mix changes with time or from animal to animal, the nature of the antiserum also changes and it may be impossible to recapture the specificity or other properties again. A second major problem with conventional immunization is that many of the most biologically interesting molecules, such as tumor or differentiation antigens, are small parts of complex mixtures. While specific antiserums can sometimes be generated by extensive and repeated absorption, the heterogeneity and unpredictability of the immune response makes it difficult to repeatedly generate large amounts of antiserum with the same properties. These and other problems with conventional immunization have discouraged and hindered the production of antiserum against many important and useful antigens and have certainly limited the use of immunoassays in the routine diagnostic laboratory to those antigens which can be obtained in relatively pure form and which induce a good immune response.
eBook - PDFClinical Biochemistry V3
Contemporary Theories and Techniques
- Herbert Spiegel(Author)
- 2012(Publication Date)
- Academic Press(Publisher)
3 Monoclonal Antibodies in Clinical Investigations PATRICK C. KUNG, TSE-WEN CHANG, AND VINCENT R. ZURAWSKI, JR. I. Introduction 89 II. Definition and Production of Monoclonal Antibody 91 III. Monoclonal Antibodies for Studying Cell Differentiation and Monitoring Diseases 93 A. Human T-Cell Development Studies 93 B. Pathology and Diagnostic Research using Anti-T-Cell Antibodies 98 C. Monoclonal Antibodies for Diagnosis of Infectious Diseases 101 D. Monoclonal Antibodies Reactive with Tumor-Associated Antigens 104 IV. Monoclonal Antibodies for Serotherapy 106 A. Organ Transplantation 106 B. Treating T- and B-Cell Lymphomas 107 V. In Vitro Treatment of Bone Marrow for Autologous Transplant 108 VI. Monoclonal Antibodies for in Vivo Radioimaging of Tumors. 109 VII. Discussion 110 VIII. Appendix: General Reviews on Perspectives, Methodology, and Applications 111 References 112 I. INTRODUCTION Throughout the history of medicine, technological breakthrough has often brought forth major advances in the diagnosis, monitoring, or thera-peutic intervention of diseases. To date, many diseases, such as cancer, autoimmune disorders, and microbial infections remain difficult to prog-nose or diagnose. In many instances ideal treatment for the disease is still 89 CLINICAL BIOCHEMISTRY Copyright © 1984 by Academic Press, Inc. Contemporary Theories and Techniques, Vol. 3 All rights of reproduction in any form reserved. ISBN 0-12-657103-1 90 Patrick C. Kung, Tse-Wen Chang, and Vincent R. Zurawski, Jr. unavailable. The recent emergence of hybridoma technology for mono-clonal antibody (MOab) production promises some solutions to these medical problems. The antibody molecule is a remarkable product of evolution. The mech-anism of its action was first postulated in the late nineteenth century.
eBook - PDFMonoclonal Antibodies
A Manual of Techniques
- Heddy Zola(Author)
- 2013(Publication Date)
- CRC Press(Publisher)
183 Chapter 7 PROSPECTS, PROBLEMS, AND LIMITATIONS IN THE USE OF Monoclonal Antibodies L SCOPE OF THE CHAPTER In the 10 years since a practical method was described for making monoclonal anti bodies, the technique has been applied in nearly every field of biology and medicine. Monoclonal Antibodies have revolutionized some of the areas in which they have been applied, by providing the reagents with which to analyze the underlying phenomena. The outstanding example is cellular immunology, which has seen the concepts of cel lular interactions in the control of immune responses develop in detail and acceptance. The basic concepts existed well before the availability of Monoclonal Antibodies, but they were based on experiments which were open to criticism because of the lack of specificity of the reagents used. Thus, allo-antisera, used in dissecting cell cooperation in mice, were contaminated with antiviral antibodies, which may or may not have affected the results. In man, experiments on cell cooperation and interaction in the immune response were based on technically difficult in vitro assays which did not re produce well in different laboratories. Monoclonal Antibodies have provided the probes for identifying and separating cells, enabling the dissection of the mixture of interacting cells which together make the ingredients of the immune response. Having provided the reagents to analyze the im mune response at the cellular level, Monoclonal Antibodies are currently being used to analyze the immune response at a further level of resolution, the molecular level. Mem brane receptors for stimulation or suppression are being analyzed and purified, and their functional sites probed with Monoclonal Antibodies. In this field monoclonal an tibodies and probes for DNA and RNA are being used as complementary tools.
eBook - PDF- Vasant V. Ranade, John B. Cannon(Authors)
- 2011(Publication Date)
- CRC Press(Publisher)
47 2 Site-Specific Drug Delivery Utilizing Monoclonal Antibodies* INTRODUCTION At the beginning of this century, Paul Ehrlich reported the discovery of antibodies. 1 Since that time, many investigators have done extensive work using a wide variety of antibody molecules in immunocytochemistry, radioimmunoassay, and clinical medi-cine. In 1976, Kohler and Milstein employed a method of somatic-cell hybridization in order to successfully generate a continuous “hybridoma” cell line capable of produc-ing monoclonal antibody (MAb) of a defined specificity. 2 Subsequently, several MAbs have exhibited specificity for target sites. It is this property of MAbs that makes them excellent candidates as carriers of therapeutic agents for delivery to specific sites. 3,4 C HEMISTR Y Antibodies are complex proteins, consisting of multiple polypeptide chains that con-tain a variety of reactive chemical groups, such as amino, carboxyl, hydroxyl, and sulfhydryl. Functionally, MAbs possess a molecular polarity based on the joining of an antigen-binding fragment (Fab) to a complement-fixing fragment (Fc). The Fab fragment is responsible for specific antigen binding, whereas the Fc fragment binds to effector cells, fixes complements, and elicits other in vivo biological responses. In order to obtain an MAb suitable for the treatment of human disease, it is neces-sary to maintain both the physical and functional properties of the antibody through-out the steps of production, isolation, purification, and modification. Antibody modification, performed to increase theoretical efficacy, can consist of conjugation of the protein to the following: radionuclides (e.g., 131I and 111In), chemotherapeutic drugs (e.g., methotrexate and vinblastine), and polypeptide toxins (e.g., ricin A chain and polkweed antiviral protein [PAP]). P OLYCLONALS VS . M ONOCLONALS Antibodies can be heterogeneous with respect to size, charge, antigen specificity, and affinity.
eBook - PDF- A.M. Campbell(Author)
- 2000(Publication Date)
- Elsevier Science(Publisher)
If, in addition, they were obliged to undertake sequential monoclonal antibody production in order to isolate a minor compon- ent of a highly antigenic mixture, the time involved and the running costs would be very extensive. If a single hybridoma line is all that is required then collaborative work with a laboratory with the appropriate facilities and experience would generally be the better course of action. More detailed costing information is given in Chapter 5. Ch. 1 GENERAL PROPERTIES AND APPLICATIONS OF Monoclonal Antibodies 17 1.3. Applications of Monoclonal Antibodies The applications of monoclonal antibody technology are generally outwith the scope of this volume and are reviewed in several others (McMichael and Fabre, 1982; Albertini and Ekins, 1981; Hammer- ling, Hammerling and Kearney, 1981; Kennett, McKearn and Bech- tol, 1980; Edwards, 1981; Yelton and Scharff, 1981). The Index Medicus lists several hundred papers each month under the heading of Monoclonal Antibodies. It is, however, possible to outline some of the major applications of monoclonal antibody technology at the present time so that the range and scope of the technique may be summarised. 1.3.1. Diagnostic uses Antibodies produced in the mouse or the rat are most commonly used for diagnostic purposes as they are more readily produced. Not only is the fusion frequency an order of magnitude higher but the animal can be hyperimmunised with the chosen antigen. The major advan- tage of Monoclonal Antibodies over conventional sera in this appli- cation is probably their ready availability for an indefinite period at a standard titre, making direct comparisons between different labora- tories comparatively simple. However, their high specificity has added greatly to the accuracy and speed of the diagnosis. Thus antibodies to common serum analytes such as protein hormones or alphafetopro- tein are already commercially marketed and are slowly replacing conventional sera.
eBook - PDFDrug Disposition and Pharmacokinetics
Principles and Applications for Medicine, Toxicology and Biotechnology
- Stephen H. Curry, Robin Whelpton(Authors)
- 2022(Publication Date)
- Wiley(Publisher)
Examples include tositumomab, ibritumomab tiuxetan, and notably, trastuzumab, which is consid- ered in this light in Box 8.1. Cancer cells are used in this way because of their propensity for rapid growth and cell division, but primarily because they are immortal and thereby provide consistency of product. 8.2 Nomenclature It can be argued that mAbs are not strictly speaking antibodies because they do not engage in an antigen– antibody interaction However, it is accepted that their nomenclature should relate to IgG and so the term anti- body is used. Monoclonal Antibodies may be referred to as being humanized, or as chimeric, depending on their constituent parts and how they have been produced. Humanized mAbs are produced from non-human species in which the amino acid sequences have been modified so that the resultant antibody is similar to pro- teins that are produced naturally in humans. The humanization technique utilizes recombinant DNA containing a gene pattern to produce the required antibody in cloned mammalian cells in a bioreactor. The term cloned is used here because a large, potentially immortal, cell line is derived from a single cell. The protein sequence of a humanized antibody is essentially identical to that of a human variant, despite the non-human origin of some of the CDR segments responsible for the ability of the antibody to bind to its target (Figure 8.2). Chimeric proteins are those produced by joining two or more genes, often from different species, to produce a fusion gene, resulting in biosynthesis of a protein with properties of both of the original proteins. In the case of human-mouse chimeric mAbs, this results in a larger sequence of mouse amino acids, but the human portion closely resembles the original human protein. Box 8.1 Stages in the production of a monoclonal antibody for use against human disease.
eBook - PDF- P. Tijssen(Author)
- 1985(Publication Date)
- Elsevier Science(Publisher)
The specificity of monoclonal anti- bodies may sometimes prove to be not as high as expected. Some may cross-react, and this cross-reactivity cannot, in contrast to poly- clonal antisera, be removed with immunosorbents (Brodsky et al., 1979). A monoclonal antibody is unable to distinguish different anti- gens if they bear the same epitope. For example, Bundesen et al. (1 980) encountered this problem with a peptide sequence common to several hormones. Kurstak et al. (1983) emphasized problems with monoclonal reagents in virus diagnosis. Monoclonal antibody production is time consuming, particularly for weak immunogens which require many fusion experiments to Ch. 5 ANTIBODY PRODUCTION 61 obtain a producing hybrid. Excellent immunogens usually yield 5-20 producing hybrids per fusion. The number of species to produce Monoclonal Antibodies is limited. Fusion of myeloma cells with cells of other species leads to rapid segregation of chromosomes (Yarmush et al., 1980). Each monoclonal antibody may have very specific properties, quite different from the average of polyclonal Ig of the same subclass. Monoclonal Antibodies may also have biological func- tions different from the corresponding polyclonal antisera and may be much more sensitive to inactivation by freezing and thawing, changes in pH or other physical properties (Mosmann et al., 1980), important for their purification. A general comparison of Monoclonal Antibodies and polyclonal antisera is given in Table 5.2.
eBook - PDFTherapeutic Monoclonal Antibodies
From Bench to Clinic
- Zhiqiang An(Author)
- 2009(Publication Date)
- Wiley(Publisher)
Cell Immunol. 200:16–26. Xu, L., H. Yee, C. Chan, A.K. Kashyap, L. Horowitz, M. Horowitz, R.R. Bhatt, and R.A. Lerner. 2008. Combinatorial surrobody libraries. Proc. Natl. Acad. Sci. USA 105:10762 – 10767. Zeidler, R., G. Reisbach, B. Wollenberg, S. Lang, S. Chaubel, B. Schmitt, and H. Lindhofer. 1999. Simultaneous activation of T cells and accessory cells by a new class of intact bispecific antibody results in efficient tumor cell killing. J. Immunol. 163:1246 – 1252. Ziegelbauer, K., and D.R. Light. 2008. Monoclonal antibody therapeutics: Leading companies to maximize sales and market share. J. Commercial Biotechnol. 14:65–72. Zoller, M.J., and M. Smith. 1982. Oligonucleotide-directed mutagenesis using M13-derived vectors: An efficient and general procedure for the production of point mutations in any fragment of DNA. Nucleic Acids Res. 10:6487 – 6500. 50 THERAPEUTIC Monoclonal Antibodies: PAST, PRESENT, AND FUTURE & CHAPTER 2 Antibody Molecular Structure ROBYN L. STANFIELD and IAN A. WILSON 2.1 Introduction 51 2.2 General Structural Features 52 2.3 Canonical Conformations 56 2.4 Fab Conformational Changes 56 2.5 Human Anti-HIV-1 Antibodies 58 2.6 Shark and Camel Antibodies 61 2.7 Summary 63 Acknowledgments 63 References 63 ABSTRACT The structural features of antibodies have been studied extensively over the years by many techniques, including electron microscopy, x-ray crystallography, and NMR. Consequently, a wealth of structural information is available for antibodies, alone and in complex with antigens ranging in size from small haptens to whole viruses. The knowledge gained from these studies has greatly facilitated the engin- eering of antibodies for use as human therapeutics. 2.1 INTRODUCTION Antibodies are the key component of the humoral adaptive immune response against foreign patho- gens. Their enormous sequence and structural diversity allows for recognition of any foreign antigen imaginable, with high affinity and specificity.
eBook - PDFMonoclonal Antibody and Immunosensor Technology
The production and application of rodent and human monoclonal antibodies
- A.M. Campbell(Author)
- 1991(Publication Date)
- Elsevier Science(Publisher)
38 MONOCLONAL ANTIBODY AND IMMUNOSENSOR TECHNOLOGY Ch. 1 GENERAL PROPERTIES AND APPLICATIONS OF Monoclonal Antibodies 39 type attitudes were not experimentally sustainable. Finally, an antigen-Mab reaction and anti-idiotype-Mab reaction for the same Mab and antigen were shown to have unrelated recognition patterns (Bentley et al., 1990). Many a&i-idiotype observations remain to be resolved. Jerne shared the Nobel prize with Kohler and Milstein in 1984 for his theo- retical description of a potential idiotypic network, but his contribu- tions to immunology extend far beyond that. Personalities apart, ac- cumulating evidence suggests that any specific mirror image of antigen and 2nd (anti-idiotype) antibody is, at least in monoclonal terms, serendipitous. On a practical point, while it is simple to obtain low-affinity polyspecific IgM Mabs from a mouse immunised with a mouse Mab, it is exceedingly difficult to obtain a high-affinity, non- cross-reactive IgG Mab. More rigorous experimental systems will be required to prove that the anti-idiotype approach is anything but a minor, low-affinity ripple, in the immune system. 1.14. Mabs as catalysts (abzymes) This application of Mabs was first described in 1986 (Tramontano et al, 1986; Pollack et al., 1986) and the field has grown rapidly since (Reviewed by Shokat and Schultz, 1990; technical descriptions by Tramontano and Schloeder, 1989 and Pollack et al., 1989). The basic concept behind catalytic Mabs is that a Mab generated to a molecule or molecules which mimic the transition state between substrate and product may accelerate the reaction by forcing the substrate into a suitable conformation for conversion into product (Fig. 1.12). Where bimolecular reactions are involved, the antibody can bring the two molecules together to significantly increase their probability of inter- 4- Fig.
- Mark Wick(Author)
- 1988(Publication Date)
- CRC Press(Publisher)
An understanding of the concept of MoAb (and an appreciation of why it has been sought like an immunologic Holy Grail) requires an appreciation of the immune system's prime imperative, variability. Hence, this section of Chapter 1 briefly outlines the biochemical and genetic basis of Ig variability and how that normative variability can be sub-verted. With this as a background, the present and future technology of MoAb production is briefly reviewed (with special emphasis on aspects of interest to the end user). No attempt is made to provide a practical hand-book for methods of MoAb production or to exhaustively review MoAb applications. Many excellent publications exist (see Refs. 12-16, for example). To a great extent this chapter is a review of reviews that may provide a con-venient starting place for the individual interested in the immunotechnology of MoAb production and also provide some caveats regarding their use. Monoclonal Antibodies: The Concept Molecular Variability of Immunoglobulins Background. Historically, variability has been recognized as a hallmark of the humoral immune response. By the 1940s, Landsteiner and his many associates had recognized that immunization of animals with even highly defined simple antigens yielded a bewildering array of antibodies [1]. Theoretical and Technical Considerations 3 Antigens that were structurally similar to the immunizing antigen could react with, or absorb, part of the antibody activity, residual activity still being detectable using the immunizing antigen. Such cross-reacting antigens showed a degree of reactivity that was roughly proportional to their degree of phylo-genetic relatedness (and hence structural similarity) to the immunizing antigen. This serological variability in response was shown to be mirrored by the bio-chemical heterogeneity of antibody molecules. Antibody activity in serum showed a considerable range of electrophoretic mobility and, to a lesser degree, molecular weight.
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