This volume contains detailed, comprehensive advice on rat, mouse and human hybridoma production. It begins with a general introduction, then describes the practical applications of the technology with photographs and protocols for everything from animal dissection to epitope analysis of antigens.

This volume contains detailed, comprehensive advice on rat, mouse and human hybridoma production. It begins with a general introduction, then describes the practical applications of the technology with photographs and protocols for everything from animal dissection to epitope analysis of antigens.

This highly practical book, and successor to Volume 13 in the Laboratory Techniques series, explores further and provides more comprehensive, autoritative information on the production of Mabs. Much new and illuminating material has been included covering the concepts behind the application of recombinant DNA technology and biosensor technology to monoclonal antibodies, and all the human Mab technology facilitated by PCR of antibody genes.Also included in this latest volume is a section focussing on other methods of obtaining B cell clones such as short-term culture and oncogene transformation and an interesting section on Mab patents.

Soon after Kohler and Milstein described the use of somatic cell hybridization for the production of murine monoclonal antibodies of desired specificity, this relatively simple technique became widely applied. Indeed, production of murine monoclonal antibodies is now considered routine by immunologists and nonimmunologists alike. However, as heterologous proteins, mouse monoclonal antibodies have one major limitation: they are immunogenic in man and, hence, their use in vivo is severely limited. An obvious solution to this problem is to produce human hybridomas with the same techniques used for the production of rodent hybrids. Unfortunately, the history of human hybridomas has been marked by substantive and often exasperating tech nical problems, and the first reports of hybrids secreting human immu noglobulin of desired specificity did not appear until 1980. These reports were met with initial enthusiasm, but it soon became apparent that while human lymphocytes might be fused, their frequency, level of Ig synthesis, and stability were such that production of human antibodies with this method was neither routine nor practical. Nonetheless, a sufficient number of investiga tors persevered, and during the next 5 years relatively efficient B-cell fusion partners as well as improved methods of Epstein-Barr virus transformation were developed. Generation of human T -T hybrids has also been achieved, although problems of chromosomal stability remain a substantial obstacle, more so than with B-cell lines.

On August 7, 1975, Kohler and Milstein published in Nature (256:495) a report describing "Continuous cultures of fused cells secreting antibody of predefined specificity. " Their report has become a classic and has already had a profound effect on basic and applied research in biology and medicine. By the time the first Workshop on Lymphocyte Hybridomas (Current Topics in Microbiology and Im munology 81, 1978) was held on April 3-5, 1978, in Bethesda, Maryland, investi gators from many laboratories had made hybrids between plasmacytomas and spleen cells from immunized animals and had obtained monoclonal antibodies reacting with a broad variety of antigenic determinants. At the time Kohler and Milstein introduced this new technology, the editors of this volume were involved in the production of antisera against differentiation antigens (K. B. B. ), histocompatibility antigens (T. ]. McK. ), and human tumor associated antigens (R. H. K. ). Because of the potential usefulness of monoclonal antibodies in these areas, we each began production of hybridomas and analysis of the resulting monoclonal reagents. One of the most interesting aspects of participation in the early stages of the development and application of hybrid oma technology has been observing how the implications of the initial observa tions gradually spread first among the practitioners of immunology and immu nogenetics, and then to other areas of the biological sciences, such as developmental biology, biochemistry, human genetics, and cell and tumor biology.