Affinity purification of tetanus toxin using polyclonal and monoclonal antibody immunoadsorbents.
Tetanus toxin has been immunopurified on immunoadsorbent columns derived from equine polyclonal antitoxin coupled to cyanogen bromide-activated Sepharose CL4B. Desorption of bound toxin in active form was achieved only when the immunoadsorbent was mixed with Sephadex G15 and this mixture overlaid on a further volume of Sephadex G15. With equine antibody, 64% of adsorbed toxin was recovered with a specific activity of 2400 limiting flocculation units (Lf)/mg protein N (1.2 X 10(8) minimum lethal doses (MLD)/mg protein N). Similarly prepared immunoadsorbent derived from murine monoclonal antitoxin of low affinity had improved desorption with less acidic desorbents, without the requirement for Sephadex G15; greater than 80% of adsorbed toxin was recovered with a specific activity of 3000 Lf/mg protein N (1.6 X 10(8) MLD/mg protein N).
Sheppard
A J
AJ
Hughes
M
M
Stephen
J
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000936:Antigen-Antibody Complex / Q000276:immunology*; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D006825:Hybridomas; D007106:Immune Sera / Q000276:immunology; D007123:Immunoelectrophoresis, Two-Dimensional; D007163:Immunosorbent Techniques; D007164:Immunosorbents / Q000276:immunology*; D011863:Radioimmunoassay; D013744:Tetanus Toxin / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D007106:Immune Sera; D007164:Immunosorbents; D013744:Tetanus Toxin