Use of SEC in the Analysis of Plasmid DNA

Analysis of plasmid DNA using SEC

Abstract

Here we present data, for different DNA plasmids analyzed using SEC, LS, RI and V. The TDA detector provides information about the DNA plasmids, including molecular weight, which cannot be analyzed using techniques such MS due to the large size of the plasmids.

Introduction

Within the biopharmaceutical industry, recombinant plasmid DNAs are used as both raw materials and active ingredients in DNA vaccines. GSK is developing several plasmid DNA vaccines, characterised using different methods. Anion exchange-HPLC (AEC) and agarose-gel-electrophoresis (AGE) are used to determine plasmid topology. Light scattering size techniques are used to provide information on molecular weight. Here we present data, for different DNA plasmids analysed using SEC, LS, RI and V. The TDA detector provides information about the DNA plasmids, including molecular weight, which cannot be analysed using techniques such MS due to the large size of the plasmids.

Method

The Model 302 TDA was coupled to a Waters Alliance 2690 HPLC. The Waters HPLC UV detector was disconnected, so that the sample flowed from the HPLC into the Model 302 detector. The TDA detectors are connected in series with UV first, LS (90 and 7°), RI then Viscometer.