Virus Purification Process Using Tangential Flow Filtration
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Solution Overview
Problem
Current methods for purifying viruses are limited in scalability and compliance with cGMP regulations, particularly in producing highly purified viruses and antigens on a commercial scale, leading to challenges in vaccine creation and clinical applications.
Innovation Solution
A multi-set process involving virus harvesting, clarification, concentration using filtration devices, and multiple separation procedures including ion-exchange and multi-modal chromatography to separate host cell contaminants and residual impurities, ensuring high purity and scalability of virus and antigen production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If high-speed ultracentrifugation is used to purify viruses, then virus purity is improved, but production quantity is limited to small quantities only
Solution Approach 1:
The purification process is divided into multiple sequential steps: initial clarification, concentration step, intermediate purification, and final polishing step. Each step targets specific impurities and progressively enhances purity while maintaining scalability for industrial quantities
Solution Approach 2:
Tangential flow filtration is introduced as an intermediary concentration step between clarification and purification. This step concentrates the virus preparation before subsequent purification steps, enabling the process to handle industrial quantities while achieving high purity
2Manufacturing precision
If density gradient ultracentrifugation is used to enhance virus purity, then virus purity is improved, but production time increases to 2-3 days and quantity is limited to micrograms to milligrams
Solution Approach 1:
The patent extracts and removes the time-consuming density gradient ultracentrifugation step from the purification process. Instead, it uses tangential flow filtration for concentration and affinity chromatography for purification, achieving comparable or superior purity in significantly reduced time
Solution Approach 2:
The patent changes the purification parameters by replacing ultracentrifugation-based methods with chromatography-based methods. This substitution maintains high purity outcomes while dramatically reducing process time and enabling industrial-scale production
3Productivity
If crude cell lysate method is used for virus preparation, then production quantity is improved, but contamination with non-virus factors increases
Solution Approach 1:
The patent performs preliminary concentration of the crude virus preparation using tangential flow filtration before purification steps. This preliminary action removes excess buffer and concentrates the virus, making subsequent purification steps more efficient and effective at removing contaminants
Solution Approach 2:
The patent implements a continuous multi-step purification process where each step builds on the previous one. The sequence of concentration, purification, and polishing steps continuously removes different classes of contaminants, progressively enhancing purity while maintaining industrial production quantities
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The process achieves highly purified viruses and antigens on a commercial scale, compliant with cGMP regulations, enabling effective vaccine production and clinical applications.
Implementation Method 1
concentrating the separated and clarified virus performed with a filtration device comprising a membrane with pores of a size not to exceed a predetermined limit
Implementation Method 2
at least one separation procedure includes ion-exchange chromatography to separate host cell contaminants from the virus
Implementation Method 3
at least one separation procedure includes a multi-modal chromatography to separate residual impurities from the virus on the basis of at least size differences between the virus and the impurities, and chemical interaction occurring between the impurities and one or more chromatography ligands
Data Source
AI summary
Various embodiments disclosed herein include methods and exemplary compositions associated with conjugation of virus and proteins (e.g., antigen) to form vaccines for delivery of immunological and other therapeutic agents, exemplary aspects of which may include harvesting viral and antigenic substances from source organisms; purifying the viral and protein (e.g. antigenic, including influenza hemagglutinin antigens) substances through methodologies described in an exemplary manner, as well as their concentration and collection; and a conjugation platform providing activation of the virus at a pH that increases binding rate and binding propensity between the virus and the protein, wherein embodiments related to the conjugation platform include controlling the ratio of virus to protein.