Cell hybridization, hybridomas, and human hybridomas.
Cell hybridization is one of the most basic cytotechnologies. The hemagglutinating virus of Japan was first used to cause cell fusion; however, polyethylene glycol is widely used now because of simplicity of procedure. This chapter first explains the principles of cell hybridization methods and then describes the practical protocols for preparing mouse hybridomas using polyethylene glycol. So far, lack of an excellent human fusion partner cell line that has high fusion efficiencies and does not produce immunoglobulin has hindered the spread of human-human hybridoma preparation methods. In the authors' laboratory NAT-30 and HO-323, human parent cell lines with high fusion efficiencies, have been established to prepare many hybridoma cell lines producing cancer-specific human monoclonal antibodies. Because NAT-30 and HO-323 cell lines are IgM producers, it is difficult to obtain IgG-producing hybridomas because the types of immunoglobulin produced by hybridomas are strongly affected by the characteristics of parent cells. Thus a nonimmunoglobulin-producing human parent cell line, A4H12, derived from human T lymphoma was established that can efficiently obtain IgG-producing human hybridomas. Another problem with preparing human hybridomas is that it is difficult to obtain B lymphocytes immunized with optional antigens for ethical reasons. To overcome this problem, in vitro immunization methods have been developed that allow exposure of a large number of B lymphocytes to cultured cancer cell or soluble antigens. The section on human hybridomas explains human fusion partners, in vitro immunization methods, and the preparation of human-human hybridomas using an electrofusion method. Finally, the application of human monoclonal antibodies to medical uses and the preparation of supranatural monoclonal antibodies are reviewed. These include multifunctional monoclonal antibodies and altered monoclonal antibodies having increased affinity and specificity by exchanging or modifying light chains.
Shirahata
S
S
Laboratory of Cellular Regulation Technology, Graduate School of Genetic Resources Technology, Kyushu University, Fukuoka, Japan.
Katakura
Y
Y
Teruya
K
K
D000818:Animals; D000911:Antibodies, Monoclonal; D006801:Humans; D006822:Hybrid Cells; D006825:Hybridomas; D051379:Mice
D000911:Antibodies, Monoclonal