Abstract
The rapid advance of phosphorothioate oligonucleotides in clinical development has accelerated the need for accurate quantitation of these compounds for quality control and bioanalytical applications (1). Whereas slab-gel polyacrylamide gel electrophoresis has been in routine use for the analysis of oligonucleotides, it has been replaced by capillary gel electrophoresis (CGE) as the technique of choice for length-based separation of synthetic oligonucleotides (2–10). This can be attributed to many factors including, superior resolution as a result of 10–100-fold higher electric fields, on line detection, and automation.
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References
Uhlmann E. and Peyman A. (1990) Antisense oligonucleotides: a new therapeutic principle. Chem. Rev. 90, 543.
Paulus A. and Ohms J. I. (1990) Analysis of oligonucleotides by capillary gel electro-phoresis. J. Chromatogr. 507, 113–123.
Guttman A., Cohen A. S., Heiger D. N., and Karger B. L. (1990) Analytical and micropreparative ultrahigh resolution of oligonucleotides by polyacrlamide gel high-performance capillary electrophoresis. Anal. Chem. 62, 137–141.
Heiger D. N., Cohen A. S., and Karger B. L. (1990) Separation of DNA restriction fragments by high performance capillary electrophoresis with low and zero crosslinked poly-acrylamide using continuous and pulsed electric fields. J. Chromatogr. 516, 33–48.
Cohen A. S., Najarian D. R., and Karger B. L. (1990) Separation and analysis of DNA sequence reaction products by capillary gel electrophoresis. J. Chromatogr. 516, 49–60.
Cohen A. S., Najarian D. R., Paulus A., Guttman A., Smith J. A., and Karger B. L. (1988) Rapid separation and purification of oligonucleotides by high-performance capillary gel electrophoresis. Proc. Natl. Acad. Sci. USA 85, 9660–9663.
Cohen A.S., Vilenchik M., Dudley J. L., Gemborys M. W., and Bourque A. J. (1993) High-performance liquid chromatography and capillary gel electrophoresis as applied to antisense DNA. J. Chromatogr. 638, 293–301.
Warren W. J. and Vella G. (1993) Analysis of synthetic oligodeoxyribonucleotides by capillary gel electrophoresis and anion-exchange HPLC. BioTechniques 14, 598–606.
United States Pharmacopoeia, XXII (1990) 1711.
Huang X., Gordon M. J., and Zare R. N. (1988) Bias in quantitative capillary zone electrophoresis caused by electrokinetic sample injection. Anal. Chem. 60, 375–377.
Srivatsa G. S., Klopchin P., Batt M., Feldman M., Carlson R. H., and Cole D. L. (1997) Selectivity of anion exchange chromatography and capillary gel electrophoresis for the analysis of phosphorothioate oligonucleotides. J. Pharm. Biomed. Anal. 16, 619-603.
Srivatsa G. S., Batt M., Schuette J., Carlson R., Fitchett J., Lee C., and Cole D. L. (1994) Quantitative capillary gel electrophoresis assay of phosphorothioate oligonucleotides in pharmaceutical formulations. J. Chromatogr. 680, 469–477.
Pearce M. and Watson N. (2001) Quality control of nucleotides and primers for PCR, in Capillary Electrophoresis of Nucleic Acids, Vol. 1 (Mitchelson K. R. and Cheng J., eds.), Humana Press, Totowa, NJ, pp. 347–352.
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Srivatsa, G.S., Pourmand, R., Winters, S. (2001). Use of Capillary Electrophoresis for Concentration Analysis of Phosphorothioate Oligonucleotides. In: Mitchelson, K.R., Cheng, J. (eds) Capillary Electrophoresis of Nucleic Acids. Methods in Molecular Biology, vol 162. Humana Press. https://doi.org/10.1385/1-59259-055-1:371
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DOI: https://doi.org/10.1385/1-59259-055-1:371
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