Quantitation of phosphorothioate oligonucleotides in human plasma

被引:130
作者
Leeds, JM
Graham, MJ
Truong, L
Cummins, LL
机构
[1] Isis Pharmaceuticals, Inc., Carlsbad
[2] Isis Pharmaceuticals, Inc., Carlsbad, CA 92008
关键词
D O I
10.1006/abio.1996.0088
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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. (C) 1996 Academic Press, Inc.
引用
收藏
页码:36 / 43
页数:8
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