Different ways to modify monoclonal antibodies.
In this paper we summarize experiments which were undertaken to create altered antibody molecules. Three different approaches were used. Established hybridoma lines were re-hybridized to mouse spleen cells to generate arrays of secondary hybridomas which express one particular heavy chain and one specificity together with a multitude of different light chains. In such hybrids the influence of light chains to the antibody combining site and the influence of affinity to antibody effector functions can be studied. Another way to obtain altered antibodies was the selection of cells producing less lytic IgM. With this technique we obtained (among many other variants) a series of mu-deletion products which were used to map the fine specificity of rat anti-mouse mu monoclonal antibodies. Both the anti-mu antibodies and the deletion variants were used to assign the Clq binding to the fourth C mu-domain demonstrating the power of mutant IgM in the structure-function analysis. In a third series of experiments we show the feasibility of generating new antibody combining sites by the methods of molecular genetics. The variable region gene of a heavy chain was placed in front of a kappa-constant region gene. The plasmid construct was transferred into mouse myeloma lines which stably express a variable heavy-constant light chain protein. Upon fusion with a light chain producing line, chimaeric light chain dimers with a functional antibody combining site were secreted. These experiments demonstrate that new series of man-made antibody molecules can be made in the future.