Restricted Access RP Chromatography for Large Polynucleotide Separation
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Traditional chromatographic methods using large pore sorbents capture both polynucleotides and impurities/excipients, making separation and analysis of large polynucleotides, such as mRNA, difficult due to slow diffusion and interference from process-related by-products and excipients.
Innovation Solution
Employing a chromatographic system with a sorbent having a pore size that substantially excludes polynucleotides, allowing them to interact only at the surface while trapping impurities and excipients within the pores, using reversed phase chromatography conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If large pore sorbents are used to separate polynucleotides, then resolution of polynucleotides is improved, but both polynucleotides and impurities/excipients are captured by the pores creating problems with isolation and analysis
Solution Approach 1:
The patent employs porous sorbent materials with specifically controlled pore sizes (e.g., 30-100 Å) that create size-based exclusion effects. The porous structure allows small molecule impurities and excipients to access the pore interior while excluding larger polynucleotide molecules, thereby achieving separation based on molecular size differences without capturing the target polynucleotides
Solution Approach 2:
The patent applies different pore size characteristics to different regions of the separation system by selecting sorbents with pore sizes matched to the specific size range of polynucleotides being analyzed. This localized optimization of pore dimensions ensures that the sorbent surface and pore structure provide appropriate interaction characteristics for separating polynucleotides from contaminants of different sizes
2Speed
If pore size is increased to improve polynucleotide separation, then mass transport rate improves, but separation of polynucleotides from process-related by-products and excipients becomes difficult
Solution Approach 1:
The patent optimizes the pore size parameter of the sorbent material to a specific range (30-100 Å) that balances mass transport kinetics with separation selectivity. This parameter optimization ensures sufficiently fast mass transport rates for efficient analysis while maintaining the size exclusion necessary to separate polynucleotides from smaller contaminants
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
Achieves sharp peaks and efficient separation of large polynucleotides from impurities and excipients, improving analysis and purification without interference, particularly suitable for mRNA and other large polynucleotides.
Implementation Method 1
restricted access reversed phase chromatography
Implementation Method 2
the average pore diameter of the porous sorbent is selected such that the polynucleotides are substantially excluded from the sorbent pores
Implementation Method 3
flowing the sample through the chromatographic system under reversed phase chromatography conditions
Data Source
AI summary
The present disclosure discusses a method of separating and/or purifying polynucleotides. The method includes injecting a sample into a chromatographic column that is packed with a porous sorbent having a pore size that substantially excludes the polynucleotides from the sorbent. This restricted access to the sorbent allows separation of large polynucleotides from each other and from smaller molecular weight impurities.