Abstract
The trace characterization of physiologically active substances with a low molecular weight (e. g., steroids, catecholamines and prostaglandins) is an important subject in clinical analysis, and specific a ntibodies against such“haptens” work as an essential analytical reagent. However, conventional antisera contain various antibody species derived from different B cell clones, whose nature prevent the constant supply of an antibody reagent with a defined quality. The hybridoma technology that was invented in 1975 by Köhler and Milstein principally resolved these problems; the cell fusion of immune spleen cells and myeloma generated the hybridoma clones, each secreting monoclonal antibody molecules with a defined and unique amino acid sequence and binding property. However, it is still not very easy to establish practical monoclonal anti-hapten antibodies, because this establishment requires an adequate cell culture, a reasonable design of hapten-carrier conjugate (immunogen), an effective immunization, an efficient antibody screening in hybridoma microcultures, etc. We succeeded in the generation of monoclonal antibodies against various haptens including steroids, endocrinedisrupting chemicals and drugs, which have been useful for developing practical immunoassays or immunoaffinity extraction systems. This article summarizes our biomedical analytical studies highlighting the technical aspect of the monoclonal antibody generation. Cloning of the antibody variable region genes and antibody engineering by gene manipulation are also discussed.
