Assessment of direct versus indirect magnetic bead-based T-cell isolation procedures followed by magnetic bead-based DNA isolation

Anna Rosenbaum; Ellen Bleck; Matthias Schneider; Georg Pongratz; Stefan Vordenbäumen

doi:10.1136/lupus-2016-000167

Assessment of direct versus indirect magnetic bead-based T-cell isolation procedures followed by magnetic bead-based DNA isolation

Lupus Sci Med. 2016 Jul 19;3(1):e000167. doi: 10.1136/lupus-2016-000167. eCollection 2016.

Authors

Anna Rosenbaum¹, Ellen Bleck¹, Matthias Schneider¹, Georg Pongratz¹, Stefan Vordenbäumen¹

Affiliation

¹ Medical Faculty , Hiller Research Center Rheumatology at University Hospital Düsseldorf, Heinrich-Heine-University , Düsseldorf , Germany.

Abstract

Objective: To compare direct and indirect bead-based T-cell isolation followed by magnetic bead-based DNA isolation.

Methods: T-cells were isolated by direct or indirect selection with magnetic bead coated antbiodies followed by magnetic bead-based automated DNA isolation in 10 healthy subjects. Purity of T-cells, purity of DNA (by A260/A280 ratio measurement) and DNA concentration were assessed.

Results: Direct and indirect labelling resulted in comparable T-cell purity (93.11±1.47% vs. 94.99±1.54%, p= 0.125) and DNA concentration per cell (50.97±14.15 ng/(mlxcell) vs. 49.53±13.62 ng/(mlxcell), p=0.492), while DNA purity was significantly higher after direct labelling (1.82±0.05 vs. 1.78±0.03, p=0.0488).

Conclusions: Both direct and indirect magnetic bead-based T-cell selection may be used prior to magnetic bead-based DNA isolation procedures.

Keywords: DNA isolation; T Cells; epigenetics; magnetic beads.