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Affinity purification of tetanus toxin using polyclonal and monoclonal antibody immunoadsorbents
Author(s) -
Sheppard A. J.,
Hughes M.,
Stephen J.
Publication year - 1987
Publication title -
journal of applied bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 0021-8847
DOI - 10.1111/j.1365-2672.1987.tb04929.x
Subject(s) - sephadex , polyclonal antibodies , chemistry , toxin , sepharose , antitoxin , monoclonal antibody , chromatography , toxoid , clostridium tetani , immunoadsorption , hapten , affinity chromatography , adsorption , microbiology and biotechnology , antibody , tetanus , biochemistry , biology , immunization , immunology , enzyme , vaccination , organic chemistry
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 × 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 × 10 8 MLD/mg protein N).