Ivancic-Jelecki Jelena, Brgles Marija, Santak Maja, Forcic Dubravko
Research and Development, Institute of Immunology Inc., Rockefellerova 10, 10 000 Zagreb, Croatia.
J Chromatogr A. 2009 Mar 27;1216(13):2717-24. doi: 10.1016/j.chroma.2008.10.087. Epub 2008 Oct 30.
Human plasma is an important medical substance and a raw material for production of various therapeutics. During blood sampling, storage and processing, genomic DNA is released into plasma from nucleated blood cells that are damaged in the course of the procedure. In order to determine the concentration of contaminating DNA in plasma, we developed a method for DNA isolation by using anion-exchange chromatography on a BIA Separations CIM (convective interaction media) diethylaminoethyl column. DNA was quantified by SYBR Green based real-time polymerase chain reaction. The concentration of cell-free, non-apoptotic DNA in plasma ranged between 0.06 and 22.5 ng/ml. As substantial volumes of plasma or whole blood are administered directly into the vascular system, a recipient is exposed to high amounts of cell-free DNA, several orders of magnitude higher than the amount found in other biologicals.
人血浆是一种重要的医用物质，也是生产各种治疗药物的原料。在血液采样、储存和处理过程中，基因组DNA会从在该过程中受损的有核血细胞释放到血浆中。为了测定血浆中污染DNA的浓度，我们开发了一种方法，通过在BIA Separations CIM（对流相互作用介质）二乙氨基乙基柱上使用阴离子交换色谱法分离DNA。通过基于SYBR Green的实时聚合酶链反应对DNA进行定量。血浆中无细胞、非凋亡DNA的浓度范围在0.06至22.5 ng/ml之间。由于大量的血浆或全血直接注入血管系统，接受者会接触到大量的无细胞DNA，其数量比在其他生物制品中发现的高出几个数量级。
