Animal product free system and process for purifying a botulinum toxin

A botulinum toxin and culture medium technology, applied in biochemical equipment and methods, enzymes, microorganisms, etc., can solve the toxicity of toxin loss of specificity, etc.

Active Publication Date: 2006-11-01
ALLERGAN INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is very difficult to dilute the toxin from milligram quantities to nanograms per milliliter, because the toxin will lose specific toxicity rapidly after the above high dilution

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] Preparation of Animal Product-Free Clostridium botulinum Seed Medium

[0089] Control seed medium was prepared using the following components per 1 liter of medium: NaCl (5 g), bactopeptone (10 g), glucose (10 g), BHI (to 1 liter), pH 8.1 (adjusted with 5N NaOH).

[0090] Prepare test (animal product-free) seed medium per 1 liter of medium: NaCl (5 g), Soy Peptone (10 g), Glucose (10 g), Hy-Soy (35 g / L, make up 1 liter of fluid medium), pH 8.1 (adjusted with 5N NaOH).

Embodiment 2

[0092] Cultivation of Clostridium botulinum in animal product-free seed media

[0093] The lyophilized Clostridium botulinum culture can be suspended in each 1ml of the control and test seed culture medium of embodiment 1, and subpackaged (every kind of seed culture medium) in two test tubes, each test tube is equipped with 10ml The corresponding seed medium is then incubated at 34°C for about 24-48 hours. 1 ml of the culture can then be used to inoculate a 125 ml DeLong Bellco flask containing 40 ml (corresponding) seed medium. Inoculated cultures can be grown in a Coy anaerobic chamber (CoyLaboratory Products Inc., Grass Lake, Mich.) at 33°C. Incubate at ±1°C for 24 hours.

Embodiment 3

[0095] Preparation of Animal Product-Free Fermentation Medium for Clostridium botulinum

[0096] Prepare basal fermentation medium using the following ingredients per 2 liters of medium: Glucose (15 g), NaCl (10 g), NaH 2 PO 4 (1g), KH 2 PO 4 (0.350g), MgSO 4 7H 2 O (0.1g), cysteine-HC (0.250g), tyrosine-HCl (0.250g), iron powder (1g), ZnCl 2 (0.250g), and MnCl 2 (0.4g).

[0097] A control fermentation medium was prepared using the following ingredients per 2 liters of prepared medium: BHI (500 ml; this corresponds to approximately 45.5 grams of dry weight bovine heart infusion), NZ-CaseTT (30 g), and basal medium (to 2 L ), pH6.8.

[0098] The basic fermentation medium can be prepared first and the pH can be adjusted to 6.8. Beef Heart Infusion (BHI) BHI was then prepared and its pH was adjusted to .8 using 5N NaOH. BHI was then added to the basal medium. The next step is to prepare NZ-CaseTT. Then add NZ-CaseTT to the basal medium that has been added with ...

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Abstract

Media and processes for the fermentation of Clostridium botulinum and obtaining a botulinum toxin for use in formulating botulinum toxin pharmaceutical compositions. The growth media can contain significantly reduced levels of meat or dairy by-products using non-animal based products to replace the animal-derived products. Preferably, the media used are substantially free of animal derived products.

Description

technical field [0001] The present invention relates to a medium and a method for obtaining biologically active botulinum toxin. In particular, the present invention relates to a substantially animal product-free medium, culture and anaerobic fermentation process for an organism, such as Clostridium botulinum, to obtain abundant, biologically active botulinum toxin. Background technique [0002] Pharmaceutical compositions suitable for administration to humans or animals for therapeutic, diagnostic, research or cosmetic purposes may comprise active ingredients. A pharmaceutical composition may also include one or more excipients, buffers, carriers, stabilizers, preservatives and / or fillers. The active ingredient in the pharmaceutical composition can be a biological product, such as botulinum toxin. Botulinum toxin can be obtained by culture, fermentation, and formulation processes that utilize one or more animal-derived preparations (eg, broth, and blood components or bloo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/64C12N1/20C12P21/02
CPCC12N1/20C12P21/02C12Y304/24069Y10S435/842C12N9/52A61P1/10A61P17/00A61P21/00A61P21/02A61P25/00A61P25/06A61P25/08A61P27/02C12P21/00
Inventor S·多诺万
Owner ALLERGAN INC
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