Oligonucleotide Purification via Tangential Flow Filtration

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Solution Overview

Problem

Current methods for purifying oligonucleotides require multiple steps and substantial operation time due to the presence of acid labile 5′hydroxy protecting groups at the 5′-O-oligonucleotide terminus, which hampers efficiency and automation in achieving high API purity, especially for chemically modified monomers or longer sequences.

Innovation Solution

The process involves the removal of the acid labile 5′hydroxy protecting group at the 5′-O-oligonucleotide terminus using tangential flow filtration with an acidic buffer solution, followed by neutralization and desalting steps, significantly reducing the number of operation steps and enhancing automation.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If multiple purification steps (reversed phase chromatography, concentration, deprotection, desalting) are used to achieve high API purity, then oligonucleotide purity is improved, but operation time and process complexity increase substantially

Engineering Contradiction:
ImproveAPI purityVSAvoidoperation time
Core Design Contradiction:
Manufacturing precisionVSLoss of time

Solution Approach 1:

The patent combines multiple separate purification steps (chromatography, concentration, deprotection, desalting) into a single integrated tangential flow filtration process. The TFF system performs concentration and deprotection simultaneously in one operation, eliminating the need for separate steps and reducing overall operation time while maintaining high API purity

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The tangential flow filtration system is designed to perform multiple functions simultaneously: it concentrates the oligonucleotide solution, removes the acid-labile protecting group through controlled acidification, and desalts the product all within a single unified process platform, thereby reducing process complexity

Inventive Principle:
Principle #6Universality (Multi-functionality)

2Manufacturing precision

If multiple purification steps are used to achieve high API purity, then oligonucleotide purity is improved, but device complexity increases

Engineering Contradiction:
ImproveAPI purityVSAvoidprocess complexity
Core Design Contradiction:
Manufacturing precisionVSDevice complexity

Solution Approach 1:

The patent consolidates multiple purification devices and operations into a single tangential flow filtration system. By integrating concentration, deprotection, and desalting functions into one TFF process, the number of separate devices and unit operations is reduced, thereby simplifying the overall device complexity

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The TFF system serves as a universal platform capable of performing multiple purification tasks through a single device configuration. The system can adjust operating parameters to achieve concentration, deprotection, and desalting without requiring separate specialized equipment for each function

Inventive Principle:
Principle #6Universality (Multi-functionality)

3Manufacturing precision

If traditional multi-step purification process is used, then adequate purity is achieved, but degree of automation is limited

Engineering Contradiction:
ImproveAPI purityVSAvoidautomation degree
Core Design Contradiction:
Manufacturing precisionVSExtent of automation

Solution Approach 1:

The patent integrates multiple manual or semi-automated steps into a single automated TFF process that can be controlled by a unified system. The combined process allows for automated parameter control, monitoring, and execution of concentration, deprotection, and desalting operations without requiring separate manual interventions for each step

Inventive Principle:
Principle #5Merging (Combining)

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

This approach reduces overall operation time, increases the degree of automation, and achieves higher purity of oligonucleotides by efficiently removing impurities, thereby improving the therapeutic application of oligonucleotides.

Implementation Method 1

removal of the acid labile 5′hydroxy protecting group at the 5′-O-oligonucleotide terminus of the oligonucleotide by way of tangential flow filtration with an acidic buffer solution

Methodology Applied
Scientific EffectTangential flow filtration: Filter (physical)

Implementation Method 2

removal of the acid labile 5′hydroxy protecting group... by way of tangential flow filtration with an acidic buffer solution

Methodology Applied
Scientific EffectHydrolysis: Hydrolysis

Data Source

PatentUS12043642B2Process for the purification of oligonucleotides
Publication Date: 2024.07.23 F HOFFMANN LA ROCHE INC

AI summary

The invention relates to a new process for the purification of oligonucleotides which comprises the removal of the acid labile 5′hydroxy protecting group at the 5′-O-oligonucleotide terminus of the oligonucleotide by way of tangential flow filtration with an acidic buffer solution. The process requires less steps and allows a higher degree of automation.