Quantitation of phosphorothioate oligonucleotides in human plasma

JM Leeds, MJ Graham, LA Truong, LL Cummins - Analytical biochemistry, 1996 - Elsevier
JM Leeds, MJ Graham, LA Truong, LL Cummins
Analytical biochemistry, 1996Elsevier
Methods are presented for the extraction of phosphorothioate oligonucleotides from human
plasma to permit quantitation by capillary gel electrophoresis. Extraction of the
phosphorothioate oligonucleotides from plasma was accomplished using two solid-phase
extraction columns, a strong anion-exchange column to remove plasma proteins and lipids,
followed by a reverse-phase column to remove salts. A second desalting step, achieved by
dialysis utilizing a membrane with a molecular weight cutoff of 2500 Da floating on distilled …
Methods are presented for the extraction of phosphorothioate oligonucleotides from human plasma to permit quantitation by capillary gel electrophoresis. Extraction of the phosphorothioate oligonucleotides from plasma was accomplished using two solid-phase extraction columns, a strong anion-exchange column to remove plasma proteins and lipids, followed by a reverse-phase column to remove salts. A second desalting step, achieved by dialysis utilizing a membrane with a molecular weight cutoff of 2500 Da floating on distilled water, was required to remove residual ionic material from the extracted sample. This method should be generally applicable to the analysis and quantitation of phosphorothioate oligonucleotides.
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