(515ap) Affnity Chromatography Elution Buffer Optimization For Production Of Ro-Ssa

Anti-(Ro) SS-A autoantibodies were described originally as precipitating autoantibodies in sera of Sjogren's Syndrome and Systemic Lupus Erythematosus patients. The (Ro) SS-A antigen is comprised or an acidic 60 kD protein that may also be associated with a RNA ranging in size from 80 to 112 bases. Extraction of Ro-SSA from bovine spleen has several steps including Immunoaffinity chromatography. Optimization of variables, such as the elution condition, is often necessary within this method. Affinity chromatography has a purification factor of 2,000- to 20,000 due to its specificity, it has limitation of dissociation of bond between biomolecules and their ligand at the cost of biological acitivity and yield.Various buffer systems utilizing different conditons such as extreme pH, ionic strength, chaotropic agents and commercially available elution buffers were investigated for high yield. Capture ELISA (Enzyme-linked Immunosorbent Assay) was perfomed to screen the optimum buffer for biological activity of Ro-SSA.