Process for enterovirus purification and inactivation and vaccine compositions obtained thereof
Inventors
Ella, Krishna Murthy • Daram, Vijaya Kumar
Assignees
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Publication Number
US-11911457-B2
Publication Date
2024-02-27
Expiration Date
Abstract
The present invention pertains to the field of industrial scale inactivation of various enteroviruses and large and industrial scale production of enterovirus vaccine compositions and combinations of various enteroviruses so obtained.
Core Innovation
The invention relates to a method for producing Enteroviral particles useful for preparation of vaccine compositions by producing Enteroviral particles in Vero cells, followed by purification of the Enteroviral particles by two step chromatography, stabilization of the purified particles, and chemical inactivation in the presence of formalin (0.025%) at 37 C for 13 days in presence of an inactivation buffer. The method includes addition of one or more stabilizers before the inactivation step and targets a recovery of the Enteroviral particles in the range of 80% to 99%.
The two step chromatography is carried out by first running size exclusion chromatography followed by ion exchange chromatography. The invention is directed to obtaining Enteroviral particles suitable for vaccine compositions.
In a related method, the invention further specifies preparation of a dose reduced inactivated Polio vaccine containing Sabin polioviruses. Purified polioviruses are stabilized by addition of M-199 medium containing glycine and tween 80, and inactivation is performed using formaldehyde 0.025% at 37 C for 13 days in presence of phosphate buffer with saline (between 2 mM to 10 mM) or sodium acetate.
Claims Coverage
The provided independent claims are clm-00001 and clm-00015. Across these claims, the inventive coverage centers on two independent features: (i) producing and purifying Enteroviral particles, including Sabin poliovirus, using a defined two-step chromatography sequence with pre-inactivation stabilization, and (ii) using specified formalin/formaldehyde inactivation conditions and buffers to achieve defined recovery and reduced losses.
Producing Enteroviral particles in Vero cells with two step chromatography, stabilization and formalin inactivation
A method for producing Enteroviral particles useful for preparation of vaccine compositions, comprising producing Enteroviral particles in Vero cells; purification of the Enteroviral particles by two step chromatography; stabilization of purified Enteroviral particles; chemical inactivation of the Enteroviral particles in presence of formalin (0.025%) at 37 C for 13 days in presence of an inactivation buffer to obtain Enteroviral particles, wherein one or more stabilizers are added before the inactivation step, recovery is 80% to 99%, and the two step chromatography is size exclusion chromatography followed by ion exchange chromatography.
Preparing a dose reduced inactivated Polio vaccine from Sabin polioviruses with stabilized formaldehyde inactivation
A method for preparing a dose reduced inactivated Polio vaccine containing Sabin polioviruses, comprising producing polioviruses in Vero cells; purification of the polioviruses by two step chromatography; stabilization of purified polioviruses by addition of M-199 medium containing glycine and tween 80; inactivation of the polio viruses by using formaldehyde 0.025% at 37 C for 13 days in presence of phosphate buffer with saline at a concentration between 2 mM to 10 mM or sodium acetate to reduce aggregation and thereby reducing poliovirus particle losses post inactivation by greater than 90% as compared to other buffers; and formulating the polio vaccine, wherein the two step chromatography is size exclusion chromatography followed by ion exchange chromatography.
The independent claims collectively cover a defined production and processing scheme for Enteroviral particles and Sabin polioviruses that combines Vero cell production, size exclusion chromatography followed by ion exchange chromatography, pre-inactivation stabilization, and chemical inactivation with formalin/formaldehyde 0.025% under specified buffered conditions, with the claims emphasizing particle recovery and reduced post-inactivation particle losses.
Stated Advantages
Recovery of the Enteroviral particles in the range of 80% to 99%.
Reducing aggregation of poliovirus particles.
Reducing poliovirus particles losses post inactivation by greater than 90% as compared to other buffers.
Documented Applications
Preparation of vaccine compositions using Enteroviral particles produced and processed as described.
Preparation of a dose reduced inactivated Polio vaccine containing Sabin polioviruses.
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