Review of Polyclonal Antibody Production Procedures in Mammals and Poultry

W. Carey Hanly, Ph.D., is an Associate Professor, Department of Microbiology and Immunology; James E. Artwohl, D.V.M., is a Clinical Veterinarian, Biologic Resources Laboratory; and B. Taylor Bennett, D.V.M., Ph.D., is an Assistant Professor and Director of the Biologic Resources Laboratory, University of Illinois at Chicago. W. Carey Hanly, Ph.D., is an Associate Professor, Department of Microbiology and Immunology; James E. Artwohl, D.V.M., is a Clinical Veterinarian, Biologic Resources Laboratory; and B. Taylor Bennett, D.V.M., Ph.D., is an Assistant Professor and Director of the Biologic Resources Laboratory, University of Illinois at Chicago. W. Carey Hanly, Ph.D., is an Associate Professor, Department of Microbiology and Immunology; James E. Artwohl, D.V.M., is a Clinical Veterinarian, Biologic Resources Laboratory; and B. Taylor Bennett, D.V.M., Ph.D., is an Assistant Professor and Director of the Biologic Resources Laboratory, University of Illinois at Chicago. In recent years the production of antibodies (Abs) in laboratory animals has become an essential part of many research projects. Investigators preparing to produce antibodies are confronted with a number of complex choices, some of which may be critical for success. The goal is to obtain high-titer, high-affinity antisera in a manner consistent with the welfare of the animals being immunized. Investigators not familiar with antibody (Ab) production require some guidelines to carry out immunizations in an appropriate manner. Recent studies of components of the immune system, their functions, and the actions of adjuvants on the immune system have yielded valuable information for understanding immune responses. The following discussion is an attempt to provide the non-immunologist with a rational basis for making decisions about choices of animal, immunization procedures, and adjuvants. An overview of the humoral immune response is included to provide a basis for understanding the function of adjuvants and immunization strategies. Additional information may be obtained from any of several immunology textbooks ( Janeway and Travers, 1994 ; Paul, 1994 ). Although optimization of procedures is still largely empirical, successful production of useful Abs can usually be accomplished by following currently available guidelines. The immune system of vertebrate species can be broadly divided into the innate system and the adaptive (acquired) system. Interactions between the two are important for the function of both. The innate system, which appears to be evolutionarily older, lacks a high level of specificity and efficiency but responds rapidly. It consists of (1) cells, such as macrophages and dendritic cells, and (2) humoral factors, such as complement. The adaptive immune system has a high level of specificity and efficiency but it responds slowly upon first encounter with a given foreign antigen (Ag). However, it often establishes long-term memory. The adaptive system can be divided into (1) the cellular immune system and (2) the humoral immune system. Adaptive cellular immunity is based in T cells (thymus-derived lymphocytes) and functions primarily to identify and dispose of infected or mutated host cells. Humoral immunity resides in the Abs present in the fluid compartments of the organism. Antibodies, made and secreted by B cells, identify extracellular foreign material within the host and help to neutralize and dispose of it. In mammals, Abs belong to one of five immunoglobulin (Ig) classes: IgM, IgG, IgA, IgD, or IgE. In avian species, they belong to one of three classes: IgM, IgY (also called IgG), or IgA ( Warr et al., 1995 ). IgG and IgA may be further subdivided into subclasses in some species. 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