Escherichia coli -derived vaccine and therapy against botulism

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Using a micro-organism to make a protein or polypeptide

Reexamination Certificate

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C435S069100

Reexamination Certificate

active

07820411

ABSTRACT:
A method of producing botulinum toxin C-terminal receptor binding domain (HCR) is disclosed. The one embodiment, the method comprises the steps of (a) preparingE. colitransformed with an expression vector comprising DNA encoding HCR protein, (b) inducing expression of the HCR protein at a reduced temperature in a culture media, and (c) purifying the HCR protein via extraction, wherein the extraction comprises a clarification by centrifugation and a filtration, wherein the purified HCR protein is at least 10 mg/L of culture medium.

REFERENCES:
patent: 5919665 (1999-07-01), Williams
Zhou et al. (Prot. Exp. Pur., 34:8-16, 2004, available online on Dec. 19, 2003).
Carstens et al. (Strategies Newsletter, 14:50-52, 2001).
Baldwin et al. (Prot. Exp. Pur., 37:187-195, Sep. 2004).
Lalli et al. (J. Cell Sci., 112:2715-2724, 1999).
Baldwin, M., et al., “The C-terminus of botulinum neurotoxin type A light chain contributes to solubility, catalysis and stability”, Protein Expr. Purif., 2004, 37:187-195.

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