A high resolution two-dimensional affinity electrophoresis has been developed, using capillary isoelectric focusing as the first electrophoresis and slab gel affinity electrophoresis as second electrophoresis. By this method 1-2 micrograms of anti-dinitrophenyl antibodies have been separated completely into several hundred homogeneous IgG spots. They are grouped into a number of families which are composed of several IgG spots of the same affinity to the hapten but of a different pI. It is suggested that each individual family is derived from one monoclonal antibody producing cell line.

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Thermodynamic analysis of the interactions of a mouse dinitrophenyl-specific myeloma protein, MOPC 3 15, with immobilized dinitrophenyl and trinitrophenyl ligands by affrnity electrophoresis* Affinity electrophoresis was applied to determine the dissociation constants and the thermodynamic parameters for the interactions between the mouse myeloma, MOPC 3 15, and trinitrophenyl (Tnp) and dinitrophenyl (Dnp) haptens. The myeloma protein consists of monomeric, dimeric, and trimeric forms of IgA. The dimer had a higher affinity to the haptens than the monomer, but its Fab' fragment had lower affinity than the monomer. All of these myeloma proteins had higher affinities to Tnp-hapten than to Dnphapten. The affinity of MOPC 3 15 proteins increased when the temperature increased. The van't Hoff plots for all ofthese interactions gave straight lines within the temperature range from 7 "C to 40 "C. The hN" were calculated by using the linear van? Hoff plots. The AS" were calculated from the AW and AGO. The AH0 and ASo were both positive. Hence, the interactions were endothermic and hydrophobic.