Monoclonal and Polyclonal Antibodies

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(Courtesy of Alex Bagley. Used with permission.) Bagley 1 Monoclonal and Polyclonal Antibodies First studied as early as 1895, antibodies comprise an integral element of the human immune response to infection. Advances in the field of immunochemistry by such pioneers as Arrhenius, Pauling, and Karl Landsteiner have provided a working model of antibody- antigen binding, while the development of X-ray crystallography has revealed the fine structural variation between the five classes of antibodies, or immunoglobulins. In the last forty years, scientists have been able to harness this knowledge to develop an array of biological assays that have since become essential in the modern molecular biology laboratory. Polyclonal antibodies bind to multiple epitopes of a given antigen and are produced by (1) immunizing a mammal – often a goat or rabbit – with an antigen, (2) bleeding the animal periodically, and (3) extracting the antibodies directly from the serum.1 The process is relatively inexpensive, and large quantities of an antibody can be isolated from a single extraction. As their name implies, monoclonal antibodies are derived from a single (‘mono-’) clone of antibody-producing B cells. After immunizing a mammal, the animal’s spleen (location of B cell production) is extracted, and the B cells are fused to immortal myeloma cells; the fused B cells and myeloma cells – or hybridomas – are screened via ELISA, and the selected hybridoma line is injected into a second mammal. The myeloma component of the hybridoma induces localized tumor growth in the animal, and an antibody-rich fluid called ascites can be extracted from the tumor; the antibodies are then isolated themselves through column chromatography.2 A powerful feature of monoclonal antibodies is that the immortal nature of the hybridoma allows for its continual reculturing, thus providing a constant source of monoclonal antibodies. Bagley 2 Diagram showing the method of production for monoclonal antibodies3: Figure removed. Please see http://ntri.tamuk.edu/monoclonal/mabcartoon.gif. Endnotes: 1 Garvey, J. S.; Cremer, N. E.; Sussdorf, D. H. Methods in Immunology; W. A. Benjamin, Inc.: Reading, Massachusetts, 1977. 2 Kohler, G.; Milstein, C. Nature 1975, 256, 495-497. 3 Natural Toxins Research Center at Texas A&M University - Kingsville. <http://ntri.tamuk.edu/monoclonal/mabcartoon.gif> Accessed November 3, 2004. .

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Perspectives on in Vitro Diagnostic Devices, Regulated by the Office Of
1 2 “This transcript appears as received from the commercial transcribing service after inclusion of minor corrections to typographical and factual errors recommended by the Technical Point of Contact.” 1 PERSPECTIVES ON IN VITRO DIAGNOSTIC DEVICES, 2 REGULATED BY THE OFFICE OF BLOOD RESEARCH AND REVIEW 3 4 FOOD AND DRUG ADMINISTRATION 5 WHITE OAK CAMPUS 6 BUILDING 31 7 10903 NEW HAMPSHIRE AVENUE 8 SILVER SPRING, MARYLAND 20903 9 10 TUESDAY, JULY 16, 2019 11 8:30 A.M. 12 13 APPEARANCES: 14 MODERATOR: TERESITA C. MERCADO, MS 15 16 WELCOME: 17 JULIA LATHROP, PHD, DETTD/OBRR 18 19 INTRODUCTION: 20 WENDY PAUL, MD, DBCD/OBRR 21 22 PRESENTERS: 23 KIMBERLY BIGLER, MLS(ASCP)CMSBB, DBCD/OBRR 1 ANNETTE RAGOSTA, MT(ASCP)SBB, DBCD/OBRR 2 ZHUGONG "JASON" LIU, PHD, DRB/DBCD 3 4 CONFERENCE 5 TUESDAY, JULY 16, 2019 6 8:30 A.M. 7 DR. LATHROP: Okay, everybody. Thank you for coming to the second day of 8 our workshop which is going to be focused on considerations for IVDs regulated by the Division 9 of Blood Components and Devices in OBRR. 10 Again, the slides will be available on the website in about two weeks, so take a 11 look for them there. And moderating today’s session is Teresita Mercado from the Division. 12 MS. MERCADO: Good morning. Welcome to session three of the IVD 13 workshop. Our first speaker will be Dr. Wendy Paul. She is the deputy director of DBCD who 14 will give us an introduction to devices in DBCD. 15 DR.
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ARP American Research Products Inc http://blog.arp1.com Monoclonal Antibodies vs Polyclonal Antibodies Author : Dan Souw Monoclonal Antibodies vs Polyclonal Antibodies: what are the differences and how they will affect your experiments Monoclonal antibodies vs Polyclonal antibodies: The different production processes Both polyclonal and monoclonal antibodies share a huge number of applications, such as IHC and WB, in common. However, the very nature of how they are produced can often make one more suitable for a given application over another. So what are the difference in their production? And how does this affect their various features? The Polyclonal Antibody Production Process In nature, the primary immune response to a foreign antigen involves the activation of large numbers of B-cells to protect the body. Each B-cell targets a single antigen, and produces antibodies against a specific epitope of that antigen. The result is the production large numbers of antibodies, each with different specificities and epitope affinities, known as ‘polyclonal antibodies’. Polyclonal antibodies can be raised in many different species, such as goats, sheep, or rabbits. To do this, the animals are typically immunized with a specific antigen to elicit an immune response. This is usually followed by further immunizations to boost the levels of circulating antibodies – the titer. Following this, the serum containing the desired antibodies is collected. This is then typically enriched using affinity purification. This process – multiple immunizations followed by affinity purification – ultimately lends itself to the production of high titer, high-affinity polyclonal antibodies against the antigen of interest. The Monoclonal Antibody Production Process Monoclonal antibodies are created with a single B-cell clone from one animal, generating 1 / 4 ARP American Research Products Inc http://blog.arp1.com antibodies to a single specific epitope.
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Antibody Production
Canadian Council on Animal Care guidelines on: antibody production This document, the CCAC guidelines on: antibody production, has been developed by the ad hoc subcommittee on immunological procedures of the Canadian Council on Animal Care (CCAC) Guidelines Committee: Dr Albert Clark, Queen's University (Chair) Dr Dean Befus, University of Alberta (Canadian Society for Immunology representative) Dr Pamela O'Hashi, University of Toronto Dr Fred Hart, Aventis Pasteur Dr Michael Schunk, Aventis Pasteur Dr Andrew Fletch, McMaster University Dr Gilly Griffin, Canadian Council on Animal Care In addition, the CCAC is grateful to the many individuals, organizations and associations that pro- vided comments on earlier drafts of this guidelines' document. In particular thanks are extended to: the Canadian Society of Immunology; the Canadian Association for Laboratory Animal Science and the Canadian Association for Laboratory Animal Medicine; Dr Terry Pearson, University of Victoria; Dr Mavanur Suresh, University of Alberta; Drs Ernest Olfert and Barry Ziola, University of Saskatchewan; and Dr Patricia Shewen, University of Guelph. © Canadian Council on Animal Care, 2002 ISBN: 0-919087-37-X Canadian Council on Animal Care 315-350 Albert Street Ottawa ON CANADA K1R 1B1 http://www.ccac.ca CCAC guidelines on: antibody production, 2002 TABLE OF CONTENTS A. PREFACE . .1 E. REFERENCES . .20 B. INTRODUCTION . .2 1. General . .20 2. Polyclonal Antibody Production . .21 C. POLYCLONAL ANTIBODY PRODUCTION 3 3. Monoclonal Antibody Production . .25 3.1 Other useful references . .27 1. Animal Selection and Care . .3 3.2 Additional useful information . .27 2. Immunization Protocol . .6 3. Standard Operating Procedures . .6 F. GLOSSARY . .27 4.
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Antigens & Antibodies II Polyclonal Antibodies Vs Monoclonal Antibodies
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