Atypical LD isoenzymes with five diffuse bands and evidence for two types of binding

We describe a plasma LD isoenzyme pattern in which all five bands are unusually diffuse and of retarded mobility, an appearance not previously reported. This was shown to be due to binding to immunoglobulin G from which the enzyme could be displaced by an antiserum to gamma chains. Protein A-Sepharose removed most of the LD activity from plasma but did not dissociate the complex, indicating that the Fab end of the Ig molecule was involved in binding. Antiserum to kappa chains displaced LD5 from the complex but had no effect on the anodal bands. By contrast NAD displaced the anodal isoenzymes but not LD5. We suggest that two types of binding are involved. The LD5 complex appears to represent a true antigen-antibody reaction whereas the binding to the faster isoenzymes does not. We also suggest that immunoprecipitation is more reliable than immunoelectrophoresis or immunodiffusion for demonstrating IgG in such complexes and may be of value in defining the nature of binding.