Purification of rabbit antispermine antiserum by affinity chromatography.
Immunoglobulins G were isolated from a crude rabbit antispermine antiserum using a protein A-Sepharose CL-4B column. Affinity chromatography coupling spermine to the Sepharose matrix was employed for separation of antispermine antibodies from these immunoglobulins. Four fractions of antispermine antibodies were isolated by stepwise elution with solutions at pH range from 4 to 1. Binding constants were determined and cross-reactivity with other polyamines was tested for each fraction of antibodies. Highly specific antispermine antibodies were found in fraction IV.
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification*; D000918:Antibody Specificity; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D002846:Chromatography, Affinity; D003429:Cross Reactions; D007074:Immunoglobulin G / Q000302:isolation & purification; D007700:Kinetics; D008722:Methods; D011073:Polyamines / Q000032:analysis; D011817:Rabbits / Q000276:immunology; D013096:Spermine / Q000276:immunology*
D000906:Antibodies; D007074:Immunoglobulin G; D011073:Polyamines; D013096:Spermine