Figure 4.

Purification of cTag8-Modified LVs in an Envelope-Independent Manner

Thawed NM LVs and cTag8 LVs pseudotyped with either RDpro, MLV-ampho, or VSV-G glycoproteins were incubated with streptavidin Dynabeads for 1 hr at room temperature. Dynabeads were immobilized by magnetic capture, and flow-through fractions were collected. Streptavidin Dynabeads were then washed 4 times with cold PBS and resuspended in cold medium in the same volume as starting viral supernatants. Viral titers (international units per milliliter) of all fractions: (1) crude (Neat), (2) re-suspended magnetic beads (Beads), and (3) post-capture incubation flow-through (Flow-Through) fractions were determined by infectivity assay on 293T cells. Data represent the viral recovery of each fraction compared to corresponding total vector input and are plotted ± SD of triplicate determinations; **p ≤ 0.01; ****p ≤ 0.0001.