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A case study: oligonucleotide purification from gram to hundred gram scale
R R Deshmukh1, K O Eriksson, P Moore
1Isis Pharmaceuticals, Inc., 2292 Faraday Ave., Carlsbad, California 92008, USA.
Nucleosides, Nucleotides & Nucleic Acids
|September 21, 2001
Summary
This study details the successful purification of a phosphorothioate oligonucleotide using anion-exchange chromatography. The process was scaled effectively, maintaining high purity and yield for therapeutic applications.
Area of Science:
- Biochemistry
- Chemical Engineering
- Pharmaceutical Sciences
Background:
- Phosphorothioate oligonucleotides are critical therapeutic agents.
- Efficient purification methods are essential for producing high-quality oligonucleotide drugs.
- Scaling up purification processes presents significant challenges.
Purpose of the Study:
- To describe the purification and scale-up of ISIS 2302, a 20-mer phosphorothioate oligonucleotide.
- To optimize anion-exchange chromatography parameters for large-scale production.
- To demonstrate the feasibility of large-scale anion-exchange purification for oligonucleotides.
Main Methods:
- Anion-exchange (AX) chromatography was employed for purification.
- Key operating parameters were optimized at the gram scale.
- The process was scaled up to hundred-gram quantities using SOURCE 30Q media.
Main Results:
- High length-based purity and yield were consistently maintained during scale-up.
- The purification process proved effective at both small and large scales.
- This represents a significant demonstration of AX purification for phosphorothioate oligonucleotides.
Conclusions:
- Anion-exchange chromatography is a viable and scalable method for purifying phosphorothioate oligonucleotides.
- Optimization of operating parameters is crucial for successful scale-up.
- The demonstrated scale-up provides a robust platform for manufacturing oligonucleotide therapeutics.