Development of Improved Method of Obtaining of Fc-fragments of Human IgM


The aim of work was to develop the improved methods of obtaining and allocating Fc-fragments of human IgM. To achieve this goal IgH hydrolysis with subsequent purification (Fс)5μ-fragments was optimized. The improved method for obtaining Fc-fragments of IgM provides papain hydrolysis of immunoglobulin in the nitrogen environment for 30 minutes allowing to achieve maximum output of Fcμ-fragments without their further degradation: isolation and purification of Fcμ-fragments by two-stage gel filtration on sephacryl S-300; control the purity of the target product in electrophoresis in polyacrylamide gel with SDS and immunodiffusion for Ouchterlony. By employing the proposed scheme, we can obtain Fcμ-fragments of high purity. Outcome of Fsμ-fragments after all stages of purification was about 15 % of the initial IgM amount in the preparation. Molecular weight of Fcμ-fragments was approximately 95 kDa.

Publication year: 
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