Status: approved; component of combination vaccines

Organizations involved:

Research Foundation for Microbial Diseases (BIKEN) – Manuf.; R&D; Tech.

Sanofi Pasteur Inc. – Manuf. other

Sanofi Pasteur S.A. – Parent

Cross ref: See the entries (#431) for Diphtheria & Tetanus Toxoids & Acellular Pertussis Vaccine Adsorbed (Tripedia), which incorporates this component vaccine; Pertussis Vaccines (#505); and Pertussis Vaccine, Acellular (#506).

Description: Acellular Pertussis Vaccine Concentrate (For Further Manufacturing Use) is an acellular Bordetella pertussis bacterial vaccine manufactured by the Research Foundation for Microbial Diseases, Osaka Univ. (BIKEN), used as a component of a combination vaccine, Diphtheria & Tetanus Toxoids & Acellular Pertussis Vaccine Adsorbed (Tripedia) manufactured by Sanofi Pasteur Inc. The vaccine concentrate contains two B. pertussis bacterial components – inactivated pertussis toxin (pertussis toxoid; PT) and filamentous hemagglutinin (FHA).

Nomenclature: Pertussis Vaccine, acellular, conc./BIKEN [BIO]; Acellular Pertussis Vaccine Concentrate (For Further Manufacturing Use) [FDA]; Pertussis Vaccine [USAN]; Pertussis Vaccine USP [USP]; whooping cough vaccine [SY]; BIKEN PT/FHA [SY]; JNIH-6 [SY]

Companies.: The vaccine concentrate is manufactured by the Research Foundation for Microbial Diseases of Osaka University (BIKEN), CBER/FDA est. no. 1156. The vaccine is sold exclusively to Sanofi Pasteur Inc., formerly Aventis Pasteur Inc., for further manufacture of DTaP vaccine (Tripedia). Aventis merged into Sanofi Aventis S.A. in late 2004.

Manufacture: Bordetella pertussis undergoes large-scale fermentation in modified Stainer-Scholte medium, with separation and purification of pertussis toxin (PT) and filamentous hemagglutinin (FHA) by salt precipitation, ultracentrifugation, and ultrafiltration. After purification, PT and FHA are combined to obtain a 1:1 ratio and treated with formaldehyde to inactivate PT (forming pertussis toxoid). The purity of the PT is assayed using sodium dodecyl sulfate polyacrylamide gel elec-trophoresis (SDS-PAGE). The vaccine concentrate is shipped to Sanofi Pasteur Inc. under controlled refrigeration.

The acellular pertussis component (JNIH-6) from BIKEN used in a large Phase III pivotal field trial in Sweden in 1986-1987 was produced by purification of PT and FHA from culture supernatant fluids of B. pertussis grown in stationary culture in Roux bottles and inactivated by exposure to formalin (formaldehyde) for 25 days. U.S. clinical trials used DTaP vaccine containing a different BIKEN acellular pertussis component mixed with Connaught’s alum-adsorbed diphtheria and tetanus toxoids. The BIKEN acellular pertussis component used in U.S. trials was manufactured in either 3,000 or 15,000 Liter fermenters and inactivated by 50 days of formalin exposure. The longer formalin incubation was used to further reduce the amount of biologically active pertussis toxin. Acellular pertussis concentrate used for the U.S. approved/marketed DTaP vaccine (Tripedia) is manufactured in 15,000 Liter fer-menter(s). Available technology (at time of approval) showed the different acellular pertussis components manufactured by BIKEN to be equivalent. See also the Tech. transfer section.

FDA class: Biologic PLA

Approvals: Date = 19920820; first approval, PLA

Indications: for further manufacturing use only by Aventis Pasteur Inc. (now Sanofi Pasteur Inc.)

Status: Besides incorporation into Sanofi Pasteur U.S. DTaP/dTap vaccines, this component is used in vaccines in Japan and, presumably, also in vaccines approved and marketed in European and other major markets worldwide.

Tech. transfer: U.S. patent 5,139,776 (a division of 4,849,358), “Method for Culturing Bordetella pertussis, a Pertussis Toxoid and a Pertussis Vaccine,” by Chazono, et al., August 18, 1992, is assigned to the Research Foundation for Microbial Diseases of Osaka University (BIKEN). Pertussis toxoid obtained by detoxifying pertussis toxin using formalin (formaldehyde) can revert to a toxic (not inactivated toxin) state during storage. The inventors found that when Bordetella pertussis is cultured in the presence of cellulose and/or a cellulose derivative, pertussis toxin (PT) and filamentous hemagglutinin (FHA) can be produced in the culture in high yield at low cost and can easily be purified from the culture. Also, the pertussis toxoid obtained by detoxifying this PT and FHA using formalin does not revert to a toxic state and the stability of the vaccine is extremely improved. The overall process for producing acellular PT and FHA pertussis vaccine comprises: culturing Bordetella pertussis in a nutrient medium containing cellulose and/or cellulose derivatives; separating the culture into a supernatant and cells of Bordetella pertussis; purification to obtain a mixed antigen comprising pertussis toxin and pertussis filamentous hemagglutinin; detoxifying the mixed antigen to obtain a pertussis toxoid; and adsorbing the toxoid on an adjuvant.