Fig. (5) Immunoaffinity chromatography of specific anti−mutacin antibody.(A) Affinity 1 ml sfGFP−column was used in chromatography to purify specific antibodies from total IgGs which were purified from the immunized rabbit by protein−A column. Total antibodies sample (5 ml) was loaded in the column (inject) and unbound fraction (flow−through) was recovered. After washing, pure anti−sfGFP IgGs were recovered from the column by elution (100mM citric acid, pH3). (B) Affinity 1 ml sfGFP−mutacin−column was used in chromatography to repurify the previous unbound fraction of IgGs to get anti−mutacin antibodies in the pure fraction. (C) ELISA test in the absence of antibodies (No Ab) or in the presence of anti−mutacin, anti−sfGFP−mutacin and anti−sfGFP (purified IgGs). Antibodies (1 mg/ml stock) were tested at dilution (1:5000), whereas, anti−mutacin was tested at dilution (1:1000) in the presence of immobilized sfGFP and sfGFP−mutacin (0.5 μg/well), mutacin (2 μg/well), or in the absence of antigens (No Ag).