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- W2189131472 abstract "Synthetic oligonucleotides are utilized for diagnostic as well therapeutic purposes (antisense drugs). Synthesis of chemically modified oligonucleotides is challenging and often results in a product of limited purity [1,2]. An LC-MS method has been developed for the characterization of oligonucleotide-based drugs and diagnostic probes [3]. The method was utilized for analysis of phosphorothioate oligonucleotides, dually labeled probes and native oligonucleotides. LC-MS is useful for nucleic acids analysis [4,5]. We used an optimized LC-MS system (see Methods) for analysis of synthetic oligonucleotides and their failure products. Phosphorothioate as well as guanidine-rich antisense oligonucleotide drugs are known to be particularly difficult to analyze [6]. Due to the chaotropic proper t ies of t r ie th ly laminehexafluoroisopropanol (TEA-HFIP) buffers, we were able to successfully separate <60mer antisense oligonucleotides [3] and identify the failure oligonucleotides in a drug form. Guidelines for column selection and optimization of mobile phase composition are discussed. The ionpair reversed-phase HPLC separation benefits from the use of 2.5 μm sorbent and elevated temperature. The separation performance was equivalent or better than ion-exchange HPLC and in some cases rivaled capillary gel electrophoresis separation. Using a 50 x 1 mm XTerra column, we were able to obtain molecular weight confirmation for ~ 1-10 pmole of oligonucleotides injected." @default.
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- W2189131472 date "2002-01-01" @default.
- W2189131472 modified "2023-09-26" @default.
- W2189131472 title "LC-MS Analysis of Therapeutic and Diagnostic Oligonucleotides" @default.
- W2189131472 hasPublicationYear "2002" @default.
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