E-Book Overview
Monoclonal Antibody Protocols provides researchers in biomedical, agricultural, and biological science with a set of detailed, easy-to-follow methods for developing and using monoclonal antibodies. The protocols emphasize techniques that optimize the outgrowth of hybridomas from primary cultures of fused cells and the use of an alternative, electric-field-mediated cell fusion technique to increase the yield of hybridomas. The book stresses antibodies produced in mice, but includes methods of producing xenogeneic hybrids that yield human, bovine, equine, and porcine monoclonal antibodies. With its detailed instructions, its comments on how to alter the various steps of a protocol in order to accommodate different materials, and its hints and tips that often make the difference between success and failure, Monoclonal Antibody Protocols provides you with a ready and indispensable source of information for preparing and using monoclonal antibodies successfully in your laboratory.
E-Book Content
CHAPTER 1
Methods of Immunization to Enhance the Immune Response to Specific Antigens InVivo in Preparation for Fusions Yielding Monoclonal Antibodies Jon A Rudbach, John L. Cantrell, and J. II Ulrich 1. Introduction The first step in preparing useful monoclonal antibodies (MAbs) is to immunize an animal with an appropriate “vaccine.” Animal and vaccine are both emphasized in the preceding sentence because this chapter describes how to generate satisfactory MAbs by maximizing interactions between the two. The term vaccine was used purposefully to connote that not only antigens of interest may be contained in the immunizing product, but carriers and adjuvants may also be included. These latter components can influence greatly the success of obtaining useful hybridomas, which produce antibodies of the desired specificity and quality. Immunization protocols for obtaining only murin