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Publication Number
US-10662412-B2
Publication Date
2020-05-26
Expiration Date
Abstract
Described herein are processes for the purification of viruses and compositions thereof.
Core Innovation
The process provides aseptic purification of virus particles by contacting a liquid medium comprising virus particles with a solid-phase matrix that has a ligand-activated core and an inactive shell. The inactive shell comprises pores having a molecular weight cut off that excludes the virus particles from entering the ligand-activated core, while a molecule smaller than the molecular weight cut off can enter the ligand-activated core.
The solid-phase matrix is added to the liquid medium as a slurry at a final concentration between 2.5% (v/v) and 30% (v/v). Purification is completed by separating the solid-phase matrix from the virus particles by filtration to produce a final virus preparation, and each step of the process is performed aseptically.
Aseptic purification is also described for Paramyxoviridae family virus particles and for measles virus particles, using the same solid-phase matrix architecture and aseptic filtration separation. The process specifies that Paramyxoviridae family virus particles and measles virus particles are greater than about 100 nm in diameter, and the contact and filtration separation produce a final Paramyxoviridae family virus preparation or a final measles virus preparation.
Claims Coverage
Independent claims cover aseptic processes for purifying virus particles using a ligand-activated core/inactive shell solid-phase matrix with pores defined by a molecular weight cut off and a filtration step to produce a final virus preparation. The inventive features focus on MWCO-based exclusion of virus entry into the core, slurry concentration, filtration separation, and aseptic performance of each step.
Aseptic solid-phase matrix purification with MWCO exclusion and filtration separation
A process of purification of virus particles comprising providing a liquid medium comprising virus particles greater than about 100 nm in diameter, contacting the virus particles with a solid-phase matrix comprising a ligand-activated core and an inactive shell comprising pores having a molecular weight cut off that excludes the virus particles from entering the ligand-activated core while a molecule smaller than the molecular weight cut off of the pores can enter the ligand-activated core, and separating the solid-phase matrix from the virus particles by filtration to produce a final virus preparation, wherein the solid-phase matrix is added as a slurry at a final concentration between 2.5% (v/v) and 30% (v/v), and wherein each step is performed aseptically.
Aseptic Paramyxoviridae purification using MWCO exclusion and filtration
A process of purification of virus particles wherein the virus belongs to the Paramyxoviridae family comprising providing a liquid medium comprising Paramyxoviridae family virus particles greater than about 100 nm in diameter, contacting the Paramyxoviridae family virus particles with a solid-phase matrix comprising a ligand-activated core and an inactive shell comprising pores having a molecular weight cut off that excludes the Paramyxoviridae family virus particles from entering the ligand-activated core while a molecule smaller than the molecular weight cut off of the pores can enter the ligand-activated core, and separating the solid-phase matrix from the Paramyxoviridae family virus particles by filtration to produce a final Paramyxoviridae family virus preparation, wherein the solid-phase matrix is added to the liquid medium as a slurry at a final concentration between 2.5% (v/v) and 30% (v/v), and wherein each step of the process is performed aseptically.
Aseptic measles virus purification using MWCO exclusion and filtration
A process of purification of virus particles of a measles virus comprising providing a liquid medium comprising measles virus particles greater than about 100 nm in diameter, contacting the measles virus particles with a solid-phase matrix comprising a ligand-activated core and an inactive shell comprising pores having a molecular weight cut off that excludes the measles virus particles from entering the ligand-activated core while a molecule smaller than the molecular weight cut off of the pores can enter the ligand-activated core, and separating the solid-phase matrix from the measles virus particles by filtration to produce a final measles virus preparation, wherein the solid-phase matrix is added to the liquid medium as a slurry at a final concentration between 2.5% (v/v) and 30% (v/v), and wherein each step of the process is performed aseptically.
The independent claims are directed to aseptic virus purification processes that use a solid-phase matrix with a ligand-activated core and an inactive porous shell defined by a pore molecular weight cut off to exclude virus particle entry, followed by aseptic filtration separation to produce a final virus preparation. The core inventive structure is the MWCO-based core exclusion and the filtration separation, with virus-size and aseptic slurry-contact conditions defining the scope for general virus particles, Paramyxoviridae family virus particles, and measles virus.
Stated Advantages
Documented Applications
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