Methods and Compositions for Increasing Toxin Production
AssigneeSanofi Pasteur Biologics, LLC
InventorsAiqi Fang, Andrew James White, Seshu Tummala, Lee Wilson
Life of this patent
Estimating the term…
Abstract
The invention provides methods and compositions (such as for example, culture media) for culturing Clostridium difficile and producing the C. difficile Toxins A and B.
Claims17
1-85. (canceled)
A medium for culturing a Clostridium difficile bacterium comprising soy peptone, yeast extract, a buffering agent, a phosphate buffer, and at least one additive selected from the group consisting of chromium trioxide, clindamycin, ascorbic acid, butyric acid, D(+)xylose, D-sorbitol, sucrose, and a combination of azaserine, adenosine, and biotin, wherein the medium is at a pH of between 6.35 and 7.45.
A bacterial culture comprising Clostridium difficile and culture medium, wherein said culture medium comprises at least one additive selected from the group consisting of chromium trioxide, clindamycin, ascorbic acid, butyric acid, D(+)xylose, D-sorbitol, sucrose, and a combination of azaserine, adenosine, and biotin.
A method of culturing Clostridium difficile comprising inoculating culture medium with Clostridium difficile, said medium comprising at least one additive selected from the group consisting of chromium trioxide, clindamycin, ascorbic acid, butyric acid, D(+)xylose, D-sorbitol, sucrose, and a combination of azaserine, adenosine, and biotin.
A method for obtaining one or more C. difficile toxins comprising the steps of:
- (a) preparing an aqueous growth medium comprising soy peptone;
- (b) inoculating the medium with a C. difficile bacterium;
- (c) culturing the inoculated medium at a pH of between 6.35 and 6.65 under conditions which facilitate growth of bacterium and toxin production;
- and
- (d) isolating the one or more C. difficile toxins from growth medium.
A method of enhancing the production from a C. difficile culture of Toxin B relative to the production of Toxin A comprising the steps of:
- (a) preparing an aqueous growth medium comprising soy peptone;
- (b) inoculating the medium with a C. difficile bacterium;
- and
- (c) culturing the inoculated medium at 37° C. to 41° C. and at a pH between pH 6.35 and pH 6.65.
Dependencies are inferred from the claim text.