IPV-DPT vaccine
The invention provides a process for producing a combined vaccine containing an inactivated Sabin strain of poliovirus, a Bordetella pertussis protective antigen, a diphtheria toxoid and a tetanus toxoid, the process including a step of producing a high-titer Sabin strain poliovirus. The inventive process for producing a combined vaccine, including a step of culturing, in the presence of from about 4 g/L to about 6 g/L of a microcarrier, Vero cells to be inoculated with a Sabin strain of poliovirus, is useful as a process for efficiently producing a combined vaccine containing an inactivated Sabin strain of poliovirus.
1. A process for producing a combined vaccine containing,
(A) type I, type II, and type III inactivated Sabin strains of poliovirus in a weight proportion of D antigens of (2-4):(80-120):(80-120),
(B) a Bordetella pertussis protective antigen,
(C) a diphtheria toxoid, and
(D) a tetanus toxoid,
the process comprising:
(a) culturing Vero cells in about 4 g/L to about 6 g/L of a microcarrier, wherein the culture medium is selected from the group consisting of ME medium, DME medium, RPMI 1640 medium and RPMI 199 medium, and wherein the culture medium contains from about 5 to about 70 vol % of calf serum or fetal bovine serum;
(b) infecting the Vero cells with type I, type II and type III Sabin strains of poliovirus;
(c) allowing the Sabin strains to proliferate;
(d) recovering a virus fluid containing the Sabin strains;
(e) inactivating the Sabin strains to produce the type I, type II, and type III inactivated Sabin strains of poliovirus of (A); and
(f) mixing the inactivated Sabin strains of poliovirus of (A with a diphtheria-tetanus-pertussis combined vaccine comprising the Bordetella pertussis protective antigen, diphtheria toxoid, and tetanus toxoid of (B)-(D).
2. The process according to claim 1 , wherein the microcarrier has a concentration of about 5 g/L.
3. The process according to claim 1 , wherein the microcarrier is a dextran microcarrier.
4. The process according to claim 1 , wherein the step of culturing the Vero cell (step (a)) is carried out on a scale of at least about 3 liters.
5. The process according to claim 1 , wherein the step of culturing the Vero cell (step (a)) is carried out on a scale of at least about 30 liters.
6. The process according to claim 1 , further comprising (d-2) a step of purifying the virus fluid.
7. The process according to claim 6 , wherein the purifying step (step (d-2)) comprises:
(i) forming the virus fluid recovered in step (d) into a pellet by ultracentrifugation,
(ii) sonicating a re-suspension of the pellet; and
(iii) purifying by column chromatography.
8. The process according to claim 7 , wherein the purification by column chromatography (iii) is carried out one time only.
9. A vaccine produced by the process according to claim 1 .
10. The process according to claim 1 , wherein the microcarrier has a concentration of about 4.5 g/L to about 5.5 g/L.