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Clinical and Diagnostic Laboratory Immunology, September 2004, p. 919-923, Vol. 11, No. 5
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.5.919-923.2004

Mass Value Assignment of Total and Subclass Immunoglobulin G in a Human Standard Anthrax Reference Serum

V. A. Semenova,* E. Steward-Clark, K. L. Stamey, T. H. Taylor Jr., D. S. Schmidt, S. K. Martin, N. Marano, and C. P. Quinn

Meningitis and Special Pathogens Branch, Division of Bacterial and Mycotic Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia

Received 19 April 2004/ Returned for modification 28 May 2004/ Accepted 22 June 2004

An anti-Anthrax Vaccine Adsorbed (anti-AVA) standard human reference serum pool, AVR414, has been prepared, and the total and protective antigen (PA)-specific immunoglobulin G (IgG) were quantified. AVR414 was prepared by plasmapheresis of healthy adults who had received a minimum of four subcutaneous injections of AVA. Mass values (in milligrams per milliliter) for total IgG and IgG subclasses 1 to 4 were determined by radial immunodiffusion. Anti-PA-specific IgG assignment (in micrograms per milliliter) was done by consensus of two complementary approaches: homologous enzyme-linked immunosorbent assay (ELISA) with affinity-purified anti-PA IgG as a calibrator and summation of mean PA-specific IgG subclass concentrations determined by IgG subclass-specific ELISA using the United States National Reference Preparation for Human Serum Proteins as a standard. The total IgG concentration assigned to AVR414 reference serum was 8.33 mg/ml. IgG subclass concentrations were the following: for IgG1, 4.48 mg/ml; for IgG2, 3.35 mg/ml; for IgG3, 0.37 mg/ml; and for IgG4, 0.30 mg/ml. The assigned mass value for total anti-PA-specific IgG was 141.2 µg/ml. Anti-PA-specific IgG subclass concentrations were the following: for IgG1, 79.6 µg/ml; for IgG2, 35.3 µg/ml; for IgG3, 3.2 µg/ml; and for IgG4, 25.3 µg/ml. Human reference serum pool AVR414 will have direct application in the standardization of anthrax serological assays, in reagent qualification, and as a standard for quantification of PA-specific IgG in humans who have been vaccinated with or otherwise exposed to Bacillus anthracis PA.

* Corresponding author. Mailing address: Microbial Pathogenesis & Immune Response Laboratory, Meningitis and Special Pathogens Branch, Division of Bacterial and Mycotic Diseases, Centers for Disease Control and Prevention, Mail Stop D-01, Atlanta, GA 30333. Phone: (404) 639-4390. Fax: (404) 639-4550. E-mail: vas7{at}cdc.gov.

Clinical and Diagnostic Laboratory Immunology, September 2004, p. 919-923, Vol. 11, No. 5
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.5.919-923.2004




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