Analysis of Oligonucleotides and their Related Substances 1st Edition by George Okafo,David Elder,Mike Webb ISBN 1906799148 978-1906799144

Chapter 1 Introduction to Oligonucleotides

George Okafo, David P. Elder and Mike Webb

1.1             What are Oligonucleotides?

1.2             Oligonucleotides as Drugs
1.3             The Discovery of the Cell Mechanism to Make Use of Double-Stranded Oligonucleotides
1.4             The Development of Oligonucleotides as Medicines
1.5             Oligonucleotide Suppliers
1.6             Quality by Design Applied to Oligonucleotide Manufacture
1.7             Regulatory Guidance
1.8             Advances in Analytical Methodology

Chapter 2 Oligonucleotide Impurities and their Origin

Hagen Cramer, Kevin J. Finn and Nanda D. Sinha

2.1             Introduction
2.2             Brief Historical Perspective of Oligonucleotide Synthesis
2.3             Raw Material Related Impurities
2.4             Process Related Impurities
2.5             Chemistry Specific Impurities  
2.6             Table of Impurities and Average Masses
2.7             Summary  
2.8             Outlook

Chapter 3             Separation of Oligonucleotides and Related Substances

Bernhard Noll and Ingo Roehl

3.1             Introduction
3.2             Chromatographic Analysis of Oligonucleotides
3.3             General Principles of Chromatographic Separation
3.4             Ion Exchange Chromatography  
3.5             Reverse Phase Chromatography  
3.6             Size Exclusion Chromatography

Chapter 4             Analytical Characterisation of Oligonucleotides by Mass Spectrometry

Patrick A. Limbach

4.1             Introduction  
4.2             MS Instrumentation
4.3             Oligonucleotides in the Gas Phase  
4.4             Method Development
4.5             Applications
4.6             Quantitative Analysis
4.7            Future Developments

Chapter 5             Analytical Characterisation of Oligonucleotides by NMR Spectroscopy

Elena Bichenkova

5.1             Introduction
5.2             Different Formats of NMR Used for Nucleic Acids  
5.3             Application of 1H NMR for Oligonucleotides
5.4             Application of 31P NMR Spectroscopy for Oligonucleotide Analogues
5.5             Diffusion Ordered Spectroscopy for Oligonucleotide Characterisation
5.6             Application of NMR for Structural Analysis of Oligonucleotides with Therapeutic or Diagnostic Potentials           
5.7             Future Perspectives for NMR Characterisation of Oligonucleotides

Chapter 6             Analytical Characterisation of Oligonucleotide using Thermal Melting Curves

George Okafo, David P. Elder and Mike Webb

6.1             Introduction
6.2             In-Silico Modelling Approaches  
6.3             Spectroscopic Methods for Determining Tm
6.4             Thermal Method for Determining Tm Values
6.5             Future Directions
6.6             Conclusions

Chapter 7             Oligonucleotide Stability and Degradation

Daren Levin

7.1             Introduction
7.2             Secondary Structure Considerations  
7.3             Types of Degradation Products
7.4             Considerations for the Analysis of Degradation Products
7.5             Storage of Oligonucleotides
7.6             siRNA Stability Case Study  
7.7             Summary