EFFECT OF SALT CONCENTRATION GRADIENT ON SEPARATION OF DIFFERENT TYPES OF SPECIFIC IMMUNOGLOBULINS BY ION-EXCHANGE CHROMATOGRAPHY ON DEAE CELLULOSE

A three-stage process, consisting of an ammonium sulfate precipitation step, dialysis desalination with microporous anion-exchange Neosepta membranes and anion-exchange chromatography on DEAE-cellulose DE-52 wa s used for the isolation of mouse monoclonal antibodies specific again st different antigens. The ascites fluids contained monoclonal antibod ies against human IgG, against horseradish peroxidase and against the heavy chain of human IgM. The effect of the salt concentration gradien t in the elution buffer was examined with the aim of optimizing chroma tographic conditions. The quality of separation of protein zones was d etermined using sodium dodecyl sulfate-polyacrylamide gel electrophore sis under non-reducing conditions. The immunoreactivity of purified mo noclonal antibodies was determined by enzyme-linked immunosorbent assa y using a solid-phase adsorbed antigens against which each monoclonal antibody type was directed. (C) 1998 Elsevier Science B.V.