Method for the production of pure virally inactivated butyrylcholinesterase

A method for producing purified, virally inactivated butyrylcholinesterase comprising:

• a. contacting a butyrylcholinesterase-containing biological fluid with a cation exchange chromatography material;
• b. contacting the biological fluid with a butyrylcholinesterase-binding affinity chromatography material;
• c. recovering butyrylcholinesterase from the affinity chromatography material;
• d. solvent-detergent treating the butyrylcholinesterase recovered from the affinity chromatography material;
• e. contacting the solvent-detergent treated product with a butyrylcholinesterase-binding affinity chromatography material;
• f. recovering butyrylcholinesterase from the affinity chromatography material;
• and
• g. pasteurizing the recovered butyrylcholinesterase.

A method for producing purified, virally inactivated butyrylcholinesterase comprising:

• a. contacting a butyrylcholinesterase-containing biological fluid with a carboxymethyl-agarose cation exchange chromatography material;
• b. concentrating the fall through from the cation exchange chromatography step by ultrafiltration;
• c. contacting the ultrafiltration concentrate with a procainamide affinity chromatography material to bind butyrylcholinesterase;
• d. eluting butyrylcholinesterase from the procainamide affinity chromatography material;
• e. solvent-detergent treating the eluate from the procainamide affinity chromatography material with about 1% (v/v) Tween-80 and about 0.3% (v/v) TnBP;
• f. contacting the solvent-detergent treated eluate with a procainamide affinity chromatography material to bind butyrylcholinesterase;
• g. eluting butyrylcholinesterase from the procainamide affinity chromatography material;
• and
• h. pasteurizing the eluted butyrylcholinesterase at about +60° C. for at least about 8 hours.

A method for producing purified, virally inactivated butyrylcholinesterase comprising:

• a. contacting a butyrylcholinesterase-containing biological fluid with a carboxymethyl-agarose cation exchange chromatography material;
• b. solvent-detergent treating the biological fluid with about 1% (v/v) Tween-80 and about 0.3% (v/v) TnBP;
• c. contacting the solvent-detergent treated fall through with a procainamide affinity chromatography material to bind butyrylcholinesterase;
• d. recovering butyrylcholinesterase from the procainamide affinity chromatography material;
• and
• e. pasteurizing the recovered butyrylcholinesterase at about +60° C. for at least about 8 hours.

A method for producing purified, virally inactivated butyrylcholinesterase comprising:

• a. contacting a butyrylcholinesterase-containing biological fluid with a cation exchange chromatography material;
• b. subjecting the biological fluid to affinity purification using a peptide ligand affinity chromatography material that binds butyrylcholinesterase;
• c. recovering butyrylcholinesterase from the peptide ligand affinity chromatography material;
• d. solvent-detergent treating the biological fluid;
• e. contacting the solvent-detergent treated biological fluid with a peptide ligand affinity chromatography material;
• f. recovering butyrylcholinesterase from the affinity chromatography material;
• and
• g. pasteurizing the recovered butyrylcholinesterase.

A method for producing purified, virally inactivated butyrylcholinesterase comprising:

• a. contacting a butyrylcholinesterase-containing biological fluid with a cation exchange chromatography material;
• b. solvent-detergent treating the biological fluid;
• c. contacting the biological fluid with at least one peptide ligand affinity chromatography material that binds butyrylcholinesterase;
• d. recovering butyrylcholinesterase from the peptide ligand affinity chromatography material;
• and
• e. pasteurizing the eluted butyrylcholinesterase.

A method for isolating butyrylcholinesterase comprising contacting a butyrylcholinesterase-containing biological fluid with a peptide ligand affinity chromatography material wherein the peptide ligand is selected from the group consisting of:

• a. alanine, lysine, aspartic acid, glutamine, isoleucine, and pro line;
• b. alanine, lysine, glycine, aspartic acid, glutamine, and asparagine;
• c. tryptophan, lysine, aspartic acid, alanine, valine, and glutamine;
• d. glycine, phenylalanine, valine, glycine, 2-naphthylalanine, and alanine;
• e. glycine, phenylalanine, histidine, glycine, 2-naphthylalanine, and isoleucine;
• f. alanine, phenylalanine, threonine, asparagine, glycine, and glutamic acid;
• g. alanine, phenylalanine, threonine, asparagine, glutamine, and glutamic acid;
• h. glycine, threonine, asparagine, tyrosine, histidine, glutamine;
• and
• i. alanine, glutamic acid, valine, aspartic acid, proline, glycine;
• and recovering butyrylcholinesterase bound to said peptide ligand chromatography material.