Preparation of highly pure toxin fragments

Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Separation or purification

Reexamination Certificate

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C530S416000, C530S417000, C530S810000, C530S811000, C436S538000, C436S532000

Reexamination Certificate

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10070764

ABSTRACT:
Toxin derivatives are prepared by proteolytic treatment of holotoxin, and their toxicity is reduced by contacting the preparation with a ligand, which can be a metal or an antibody or another ligand. This ligand selectively binds to the toxin but not to the toxin derivative. Removing the ligand and toxin bound to the ligand further reduces toxicity. A second ligand is used to remove conjugates of the toxin and the first ligand. Compositions contain the purified derivative, optionally plus the toxin and the ligand.

REFERENCES:
patent: 5601826 (1997-02-01), Halpern
patent: 40 37 809 (1992-06-01), None
patent: WO 96/12802 (1996-05-01), None
patent: 9803690 (1999-02-01), None
Rosetto et al. Biochem. J. 285: 9-12, 1992.
Hallis et al. In: Botulinum and Tetanus Neurotoxins. (Ed) DasGupta et al. Plenum Press, New York, pp. 433-436, 1993.
Gimenez, J.A. and DasGupta, B.R., “Botulinum Type A Neurotoxin Digested with Pepsin Yields 132, 97, 72, 45, 42, and 18 kD Fragments,”J. Pro. Chem. 12:351-363, Plenum Publishing Corporation (1993).
Schiavo, G. et al., “Botulinum Neurotoxins Are Zinc Proteins,”J. Biol. Chem. 267:23479-23483, The American Society for Biochemistry and Molecular Biology, Inc. (1992).
Derwent Patent Abstract. File No. 351, Accession No. 9065531, English language abstract of DE 4037809A.

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