Method for continuous purification of biological material using immunosorbent.
A mechanical device for the continuous purification of biological material using immunosorbent was developed. The system consists of heat-sealed nylon pouches containing agarose-bound antibody, attached to an endless 35 mm wide Mylar belt that passes through four chambers sequentially. The biological material is bound and dissociated, and the immobilized antibody is regenerated for repeated isolation and purification of antigen. The belt design incorporates features to minimize carry-over between chambers and prevent damage to the agarose-bound antibody in repeated passes through the system. An existing batch method for the purification of human placental alkaline phosphatase using immobilized rabbit antisera was adapted to continuous purification in the device. The belt contained a low affinity immunosorbent and made five complete passes through the system. A decrease in antigen binding capacity between free immunosorbent suspensions and belt immunosorbent in pouches was observed. This was shown to be the result of the diffusion resistance offered by the pouch and the short exposure times of each pouch in the chambers. A decrease in antigen binding capacity between successive belt passes was also observed, and resulted from the inability of the agarose in the pouches to resuspend completely after each pass. The low efficiency of the agitation method and the roller device used to squeeze the pouches were the reasons for this deficiency.
D000469:Alkaline Phosphatase / Q000276:immunology / Q000302:isolation & purification; D000936:Antigen-Antibody Complex; D000941:Antigens; D001688:Biological Products / Q000302:isolation & purification*; D011232:Chemical Precipitation; D006801:Humans; D007106:Immune Sera; D007136:Immunoglobulins; D007158:Immunologic Techniques / Q000295:instrumentation; D007164:Immunosorbents; D007700:Kinetics; D010920:Placenta / Q000201:enzymology
D000936:Antigen-Antibody Complex; D000941:Antigens; D001688:Biological Products; D007106:Immune Sera; D007136:Immunoglobulins; D007164:Immunosorbents; D000469:Alkaline Phosphatase