Quantitative capillary gel electrophoresis assay of phosphorothioate oligonucleotides in pharmaceutical formulations

Srivatsa, G. Susan; Batt, Michael; Schuette, Jodi M.; Carlson, Ronald H.; Fitchett, Jonathan R.; Lee, Claudia; Cole, Douglas L.

doi:10.1016/0021-9673(94)85145-x

Cited by 68 publications

(22 citation statements)

References 8 publications

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“…In the first attempt a conventional CGE system [18,19] (CGE Buffer-1) was tested. The performance of CGE Buffer-1 for a 5-25 base long oligodeoxythymidine-phosphorothioate (p(dT) ) is shown in Fig.…”

Section: Conventional Single-base Resolution Antisense Ce-cge Systemmentioning

confidence: 99%

Capillary gel electrophoresis of therapeutic oligonucleotides – Analysis of single‐ and double‐stranded forms

Szekeres¹,

Kiessig²,

Schwarz³

et al. 2009

Electrophoresis

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Recently, several therapeutic double-stranded (ds) oligonucleotides (ODNs) are in pharmaceutical development. During quality control, these therapeutic molecules have to be characterized with respect to their identity, their content and their impurity profile. It follows that the ds molecule as well as its process- and product-related impurities have to be quantified. The single strands are considered as process as well as product-related impurities in the ds drug substance. Applying well known, conventional, single-base resolution CE-CGE systems developed for the quality control of single-stranded antisense ODNs in the early 1990s, it turned out that the ds ODNs under investigation are migrating in broad, splitted peaks between the peaks reaction zones are observed. It follows that the quantification of the single strands in the drug substance as well as quantification of other product-related impurities, e.g. n-1; n-2 (loss of one and two bases (n), respectively) etc., are not possible without adaptation of the test system. The paper shows how the test system was adjusted in order to determine single-stranded strands as well as ds strands next to each other quantitatively in the ds drug substance under investigation.

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Section: Conventional Single-base Resolution Antisense Ce-cge Systemmentioning

confidence: 99%

Capillary gel electrophoresis of therapeutic oligonucleotides – Analysis of single‐ and double‐stranded forms

Szekeres¹,

Kiessig²,

Schwarz³

et al. 2009

Electrophoresis

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show abstract

“…Calculation of the concentration of RPI.13919 putative metabolites, RPI.14212 and RPI.14213, was achieved as previously described (20,25) …”

Section: Calculationsmentioning

confidence: 99%

“…During the past few years, capillary gel electrophoresis has emerged as a state of the art analytical technique to analyze anionic polymers such as antisense oligonucleotides (19) largely because of the inherent n Ϫ 1 resolution obtained with this method. The well-documented electrokinetic injection bias can be corrected with the use of a proper internal standard, allowing for quantitative measurements (20).…”

mentioning

confidence: 99%

Quantitative Determination of a Chemically Modified Hammerhead Ribozyme in Blood Plasma Using 96-Well Solid-Phase Extraction Coupled with High-Performance Liquid Chromatography or Capillary Gel Electrophoresis

Bellon¹,

Maloney²,

Zinnen³

et al. 2000

Analytical Biochemistry

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“…Using the data obtained from CGE analysis, quantitation of phosphorothioate oligonucleotides was carried out according to the work of Srivatsa et al (21). The amount of oligonucleotides in the samples (n ON ) is calculated by…”

Section: Calculation and Data Analysismentioning

confidence: 99%

Quantitative Analysis of Modified Antisense Oligonucleotides in Biological Fluids Using Cationic Nanoparticles for Solid-Phase Extraction

Gerster

Schewitz

Fritz

et al. 1998

Analytical Biochemistry

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Quantitative capillary gel electrophoresis assay of phosphorothioate oligonucleotides in pharmaceutical formulations

Cited by 68 publications

References 8 publications

Capillary gel electrophoresis of therapeutic oligonucleotides – Analysis of single‐ and double‐stranded forms

Capillary gel electrophoresis of therapeutic oligonucleotides – Analysis of single‐ and double‐stranded forms

Quantitative Determination of a Chemically Modified Hammerhead Ribozyme in Blood Plasma Using 96-Well Solid-Phase Extraction Coupled with High-Performance Liquid Chromatography or Capillary Gel Electrophoresis

Quantitative Analysis of Modified Antisense Oligonucleotides in Biological Fluids Using Cationic Nanoparticles for Solid-Phase Extraction

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