Human immune interferon

Claim 1

A process for purifying crude human immune interferon which comprises: (a) contacting a solution of crude interferon with Controlled Pure Glass beads for a period of time sufficient to accomplish equilibrated adsorption of the crude interferon to the beads in the presence of a neutral pH buffer, washing the glass beads having interferon adsorbed thereon with a chemically compatible buffer to remove the unbound contaminant until the optical density of the eluate at 280 nm is about 0, washing the glass beads having interferon adsorbed thereon with a neutral pH buffer solution until the optical density at 280 nm is about 0, eluting the adsorbed immune interferon from the glass beads with an ammonium sulfate solution; (b) contacting the solution of interferon with an adsorbent selected from the group consisting of Concanavalin A-Sepharose, lentil lectin-Sepharose and pea lectin-agarose for a period of time sufficient to accomplish equilibrated adsorption of the interferon to the absorbent in the presence of a neutral pH buffer, washing the adsorbent having interferon adsorbed thereon from the soluble fraction with ammonium sulfate until the optical density of the eluate at 280 nm is about 0, washing the absorbent with a neutral pH buffer until the optical density of the eluate at 280 nm is about 0, eluting the adsorbed interferon with a buffer containing a member of the group consistng of alpha-methyl D-mannoside and 1-methyl D-glucoside; (c) contacting the interferon with a member selected from the group consisting of Heparin-Sepharose or Procian Red-agarose for a period of time sufficient to accomplish equilibrated adsorption of the interferon to the member equilibrated in a neutral pH buffered solution, washing the member containing adsorbed interferon thereon with first a neutral pH buffered solution containing a sugar selected from the group consisting of alpha-methyl D-mannoside and 1-methyl D-glucoside to remove the unbound material until the optical density of the eluate at 280 nm is about 0, washing with a neutral pH buffered solution until the optical density is about 0 to remove the bound material eluting the adsorbed interferon with a second neutral pH buffered solution containing a highly concentrated salt; (d) dialyzing the partially purified interferon against a chemically compatible buffer and contacting the solution of dialyzed partially pure interferon with a column of cationic exchanger resin for a period of time sufficient to accomplish equilibrated adsorption of the interferon to the exchanger in the presence of a buffer at pH about 9.0 to 10.0, washing the resin having interferon adsorbed thereon from the soluble fraction with a buffered solution until the optical density of the eluate at 280 nm is about 0, eluting the adsorbed interferon with a desorbing buffer of 20 mM tris-HCl at pH 9.0-10.0 containing 50 mM NaCl; and (e) further purifying by subjecting the elution fraction to gel-filtration treatment equilibrated in a neutral pH buffered solution containing a highly concentrated salt, and eluting whereby the solutes within the elution fractions are separated according to their molecular weights; and collecting the solute fraction containing a near homogeneous interferon preparation.