Development of Improved Method of Isolation and Purification of Human IgE
We aim at creating and evaluating the acceptability of improved methods of isolation of high purity human IgE suitable for use in highly sensitive immuno-analytical methods. To this end, we generalize physicochemical and biological properties of immunoglobulins of different classes and analyze key methodological techniques used for isolation and purification of human IgE (ion-exchange chromatography on DEAE-cellulose, gel filtration on Sephadex G-200 and S-300 Sephacryl different options immunoaffinity chromatography). In addition, we justify circuit isolation and purification of human IgE based on own research data, as well as on other authors’ data. The technique is based on combination of the following methodological techniques: sequential removal of serum human IgG (by affinity chromatography on protein G), human IgA and IgM removal (using immunoaffinity chromatography based on sorbents with anti-IgA and anti-IgM monoclonal antibodies), allocation of human IgE by dual-gel filtration on Superdex 200. In addition, we perform the quality control of IgE by immunodiffusion for Ouhterloni and electrophoresis in polyacrylamide gel in reducing conditions. Using the proposed scheme, we obtain human IgE high purity. The human IgE output after all treatment phases is about 42 % of the initial amount of immunoglobulin E in the serum.