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CVB-DAT-0056.03 Page 1 of 2 

Controlled by: APHIS Center for Veterinary Biologics, 515-337-6100 

United States Department of Agriculture

Animal and Plant Health Inspection Service

Center for Veterinary Biologics

P. O. Box 844

Ames, IA 50010

1. Reagent Name: Clostridium novyi type B (alpha) Antitoxin

2. Strain or Source: C. novyi Type B Strain CN 234.3 was used to produce the immunizing 

antigen. 

3. Lot Number: IRP 507 (04) 

4. Fill Date: June 24, 2004 

5. Expiration Date: February 28, 2029 

6. Intended Use: IRP 507 (04) serves as the standard antitoxin when evaluating the potency 

of C. novyi alpha antitoxin. 

7. Instructions for Use: IRP 507 (04) contains 140 alpha antitoxin units per mL (AU/mL). 

A dilution of standard antitoxin containing 0.1 AU/mL is used in the alpha toxin-neutralization 

test as described in title 9, Code of Federal Regulations (9 CFR), section 113.108. The dilution 

may be prepared by adding 1.0 mL of well mixed IRP 507 (04) to 13 mL of peptone diluent 

(1.0% peptone, 0.25% sodium chloride, pH 7.2). The antitoxin is further diluted by adding 

1.0 mL of the solution containing 10 AU/mL to 9.0 mL of diluent. The final dilution, containing 

0.1 AU/mL, is prepared by adding 1.0 mL of the solution containing 1.0 AU/mL to 9.0 mL of 

diluent. The antitoxin diluted 1:14 is stable when stored at -70°C or lower. 

8. Test of Reagent: 

Determination of antitoxin titer - The antitoxin titer was determined by injecting mice 

intravenously with 0.2 mL of diluted antitoxin mixed with 0.1 L+ dose of toxin (the smallest 

amount of toxin which, when mixed with 0.1 unit of antitoxin, causes death in at least 80% of the 

animals within 72 hours) and 0.1 Lo toxin dose (the largest amount of toxin which, when mixed 

with 0.1 unit of antitoxin, causes no deaths in animals within 72 hours). The antitoxin titer of IRP 

507 (04) was confirmed by comparing the results of mice injected with toxin-antitoxin mixtures 

that contained 1.0 mL of IRP 507 (04) diluted 1:1400 to the results of mice injected with toxinantitoxin mixtures that contained 1.0 mL of C. novyi International standard antitoxin possessing 

0.1 AU/mL. 

Precautions: There are no known hazards associated with the use of this reagent.
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CVB-DAT-0056.03 Page 2 of 2 

Controlled by: APHIS Center for Veterinary Biologics, 515-337-6100 

Sterility test - The antitoxin was tested for sterility and found to be free of viable bacteria and 

fungi according to procedures outlined in 9 CFR 113.26. 

9. Container Size, Type, Weight, or Volume: Two-mL glass vials containing 1.3 mL of 

antitoxin. 

10. Storage Conditions: Store at -70°C or lower. 

11. CVB Technical Contact: Bacteriology Section, Center for Veterinary Biologics, 

(515) 337-6100 or FAX (515) 337-7673.

12. Origin and Passage History: N/A 

13. Method of Preparation: Goats with no history of clostridial vaccinations received 

multiple injections of C. novyi alpha toxoid and toxin during a 10-month period. Sera from the 

hyperimmunized goats were fractionated with ammonium sulfate and the immunoglobulin 

dialyzed against 0.01 M phosphate buffered saline, pH 7.2. The antitoxin was passed through a 

sterile Millipore filtration unit containing a 0.22-μm membrane, and mixed with 15% v/v of 

sterile glycerol. No other preservative was added. 

14. Other: None 

Reagent orders and feedback should be sent including phone number to the following email 

address: VS.DB.CVB.Reagent.Requests@usda.gov

Reagent orders forms (APHIS Form 2018) can be found on the CVB website.