انساني ALG خالص سازي آنتي لنفوسيت گلوبولين

Purification of human antilymphocyte globulins

OBJECTIVES: Anti-lymphocyte globulin (ALG) is a polyclonal anti human lymphocyte surface markers which is produced in rabbits. ALG is one of the potent immunosuppressive agents which is currently used for preventing organ allograft rejection, treatment of aplastic anemia and some autoimmune disorders. ALG causes elimination of human lymphocytes from the peripheral blood circulation, regulation of cytotoxic activities and apoptosis.The goal of this project was purification of ALG by a simple, rapid and inexpensive method, ion exchange chromatography. METHODS: ALG etched serum of immunized rabbit with human lymphocytes was diluted with phosphate-buffered saline (PBS, pH 7A) in a ratio of 1:1 and precipitated with ammonium sulfate in the final concentration of 50%.The precipitant was rewashed with 50% ammonium sulfate, dialyzed against Iris-phosphate pH=8.1 and applied to ion-exchange chromatography on DEAE-Sepharose 6B.ALG tithed fraction was eluted with starting buffer; Tris-Phosphate (pH--$.1) and final buffer; ms-Phosphate containing 50 mM NaCl (pH=8.1). RESULTS: The purity of ALG with high percent purity was confirmed by SDS-PAGE, and efficiency of it by histocompatibility assay. CONCLUSION: This preparation is comparable in purity to those of commercially available standard ALG in the benefit of self sufficiency. The 1180 dilution of purified ALG can lyse human lymphocytes properly in histocompatibility testing which verifies its high efficiency.