(419h) Facile Isolation and Recovery of Biological Molecules with Chemically Triggered Degradable Polyacryamide Gel Electrophoresis

Polyacrylamide gel electrophoresis (PAGE) has been widely used in the analysis and separation of biological molecules, including proteins, DNA and RNA. For further analysis and manipulation of separated biological molecules, it is sometimes necessary to recover them from polyacrylamide gels. Most commonly, biomolecules are isolated from polyacryamide either by diffusion or electroelution. The poor diffusion of large biomolecules makes the methods slow and low-yielding. An alternative approach that has been proposed to reduce recovery time and increase yield is to incorporate degradable crosslinkers into the gel. Although degradable electrophoresis gels have been used to isolate biological molecules with high yield, they often require very limiting, harsh conditions, such as, acidic or basic buffer systems. To solve these limitations, we have developed novel crosslinkers consisting of α-azido ether functionality which can be degraded by bio-orthogonal chemical triggers. Here, we show that large biological molecules such as plasmid DNA can be separated by PAGE containing the new crosslinker, N₃-EG₂, and liberated with high yield via a chemical trigger. In addition, we find that the degradation conditions do not affect the activity of the biological molecules. We believe that N₃-EG₂ crosslinkers could be widely applied to biological molecule recovery and combined with other technologies like proteomics.