Editing VLP Expression and Purification (section)

=== In vitro encapsulation of GFP within Qβ-VLPs ===

<br/>VLPs were expressed and purified as described above. DyLight 633-labeled VLPs (10mg/mL)<br/>were incubated in disassembly buffer (20 mM Tris-HCl, 50 mM NaCl, 6M urea and 10 mM<br/>dithiothreitol) at 4 °C for one hour to disassemble VLP into CP dimers and posteriorly dialyzed<br/>against 10 mM acetic acid with 50 mM NaCl. Dialyzed CP dimers were then applied to a<br/>Sephadex G75 column and concentrated by a centrifugal filter unit (3.5 kDa MWCO).<br/>Subsequently, DyLight 633 labeled Qβ-VLPs were combined with a five-fold molar excess of<br/>GFP in dialysis tubing (3.5 kDa MWCO). VLP dimers and GFP were dialyzed against 2L of<br/>reassembling buffer (20 mM Tris-HCl, pH 7.5, 50 mM NaCl) three times at 4 °C for at least 24<br/>h. Excess GFP and unassembled dimers were removed by centrifugation (100 kDa MWCO).