Improved methods for enterovirus inactivation, adjuvant adsorption and dose reduced vaccine compositions obtained thereof
AssigneeSerum Institute of India Private Limited
InventorsRajeev Mhalasakant DHERE, Sambhaji Shankar PISAL, Jagdish Kamalaji ZADE, Rajendra Narayan SABALE
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Abstract
The present invention is directed to improved methods of Enterovirus inactivation by formaldehyde in presence of tromethamine buffer resulting in maximum recovery of D-antigen. Subsequent adsorption of said sIPV on aluminium hydroxide provides significantly dose reduced sIPV compositions.
Claims17
A method for producing a composition comprising Enteroviral particles, wherein the method comprises the steps of:
- a) producing a medium containing the Enteroviral particles;
- b) purification of the Enteroviral particles from the medium;
- c) stabilization of purified Enteroviral particles;
- d) formalin inactivation of the Enteroviral particles whereby during at least a part of the inactivation a buffer other than a phosphate buffer is present at a concentration sufficient to prevent or reduce aggregation of the Enteroviral particles thereby reducing the D-antigen losses post inactivation by 8 to 10 fold as compared to phosphate buffer;
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- e) adsorption of Enteroviral particles on aluminum salt adjuvant whereby percentage adsorption on alum is atleast 95%.
The method according to claim 15, wherein the method for preparing the dose reduced inactivated Polio vaccine containing Salk or Sabin polioviruses comprises the steps of:
- a) producing a medium containing the polioviruses;
- b) purification of the polio viruses from the medium;
- c) stabilization of purified polioviruses by addition of M-199 medium containing glycine;
- d) inactivation of the polio viruses by using formaldehyde 0.025% at 37° C. for 5 to 13 days in presence of TRIS buffer at a concentration between 30 mM to 60 mM to prevent or reduce aggregation of the poliovirus particles thereby reducing the D-antigen losses post inactivation by 8 to 10 fold as compared to phosphate buffer;
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- e) adsorption of inactivated polioviruses on Alum hydroxide adjuvant having concentration between 2 to 2.5 mg/dose, whereby percentage adsorption on Alum hydroxide is greater than 95% for Type 1, Type 2 and Type 3.
The method according to claim 15, wherein the method for preparing the dose reduced inactivated Polio vaccine containing Salk or Sabin polioviruses comprises the steps of:
- a) producing a medium containing the polioviruses;
- b) purification of the polio viruses from the medium;
- c) stabilization of purified polioviruses by addition of M-199 medium containing glycine;
- d) inactivation of the polio viruses by using formaldehyde 0.025% at 37° C. for 5 to 13 days in presence of TRIS buffer at a concentration between 30 mM to 60 mM to prevent or reduce aggregation of the poliovirus particles thereby reducing the D-antigen losses post inactivation by 8 to 10 fold as compared to phosphate buffer;
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- e) adsorption of inactivated polioviruses on Alum hydroxide adjuvant having concentration between 2 to 2.5 mg/dose, whereby percentage adsorption on Alum hydroxide is greater than 95% for Type 1 and Type 3.
Dependencies are inferred from the claim text.