PERFUSION IMMUNOAFFINITY CHROMATOGRAPHY AND ITS APPLICATION IN ANALYSIS AND PURIFICATION OF BIOMOLECULES

Immunoaffinity cartridges have been prepared by immobilizing the monoc lonal antibody to bovine growth hormone (bGH) and bovine growth hormon e releasing factor (bGRF) on the streptavidin-coated perfusion media. The cartridge-immobilized antibody to bGH has been used for the analys is of standard sample of recombinant bGH (r-bGH). The cartridge immobi lized bGRF has been applied for purification of the C- and N-terminal antibodies and their conjugates with horseradish peroxidase. The purit y of the antibody fractions purified by bGRF cartridges has been teste d by capillary zone electrophoresis. It has been shown that the immuno affinity cartridges prepared by immobilizing the antigen offer more bi ological specificity for the purification of antibody than the Protein G cartridges. The stability and life time of the immunoaffinity cartr idges are mainly dependant on the stability of the biomolecules immobi lized on them. Immunoaffinity cartridges based on the perfusion media offer us the tools for rapid analysis and purification of antigens or antibodies as well as for determination of the biological activity bet ween them at a very low back pressure in the columns.