Immobilized Poly(N)Polymerase for Homogeneous RNA Production
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Solution Overview
Problem
Current methods for producing stable and homogeneous poly(N)RNA molecules for gene therapy and genetic vaccination are inefficient, requiring large amounts of poly(N) polymerase (PNP) and involving complex purification processes, with existing kits not suitable for industrial-scale production.
Innovation Solution
Immobilization of PNP onto a solid support allows for repeated use, enhanced stability, and simplified purification, enabling the production of homogeneous poly(N)RNA molecules through a more cost-effective and time-efficient process.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If PNP is used in solution for polynucleotidyl reaction, then the reaction can proceed, but large amounts of PNP are required and complex purification processes are needed
Solution Approach 1:
The PNP enzyme is extracted from the solution phase and immobilized onto a solid support matrix. This extraction allows the enzyme to be retained on the solid support while enabling repeated use, thereby reducing the quantity of PNP needed for each reaction and eliminating complex purification steps.
Solution Approach 2:
A solid support matrix serves as an intermediary carrier for the PNP enzyme. This intermediary allows the enzyme to be固定在 a stable platform, facilitating both repeated use and simplified separation from the reaction mixture, thus improving productivity while reducing enzyme consumption.
2Productivity
If PNP is used in solution, then the reaction can proceed, but the process is time-consuming and not suitable for industrial-scale production
Solution Approach 1:
By extracting the PNP from solution and immobilizing it on a solid support, the system enables rapid separation of the enzyme from the reaction mixture through simple filtration or decantation. This eliminates time-consuming purification steps and allows for faster processing cycles suitable for industrial production.
Solution Approach 2:
The immobilized PNP on solid support can be reused across multiple reaction cycles without requiring re-purification between uses. This continuous reuse capability eliminates repeated purification time losses and maintains high productivity over extended production periods.
3Ease of manufacture
If existing kits are used for producing poly(N)RNA, then the process is straightforward, but they are not suitable for industrial-scale production
Solution Approach 1:
The solid support matrix acts as an intermediary that maintains the simplicity of the production process while enabling industrial scalability. The solid support allows for easy handling, filtration, and reuse of the enzyme, combining operational simplicity with the capacity for large-scale production.
Solution Approach 2:
The invention changes the physical state parameter of the PNP from dissolved in solution to immobilized on solid support. This parameter change maintains ease of manufacture through simple filtration and reuse while dramatically improving scalability for industrial production by enabling enzyme recovery and repeated use across multiple batches.
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 immobilization of PNP enzymes facilitates the production of stable and homogeneous poly(N)RNA molecules, improving the efficiency and scalability of RNA-based therapeutics while reducing chemical and material usage.
Implementation Method 1
a poly(N)polymerase (PNP) which is immobilized... for producing polynucleotidylated ribonucleic acid (poly(N)RNA) molecules
Implementation Method 2
Immobilization of PNP onto a solid support allows for repeated use, enhanced stability, and simplified purification
Data Source
AI summary
The present invention relates to an immobilized poly(N)polymerase (PNP), methods of producing said PNP and uses thereof. Further disclosed is an enzyme reactor and kit comprising the PNP for producing polynucleotidylated ribonucleic acid poly(N)RNA)molecules which are useful in gene therapy, immunotherapy, protein replacement therapy and/or vaccination.