Figure 8. Analysis of Protein Interactions with NTA-Modified and Control Liposomes.

Liposomes containing either 1% tris-NTA-DAP or 1.5% POPG with 5% mPEG were recovered from the plasma of mice after intravenous injection. Each lane represents recovery of liposomes from an individual mouse, which accounts for the variable amount of total protein per well. Liposomes were purified to remove unassociated proteins and run on a 4–15% Tris-Glycine SDS Page Gel. Circles (●) denote proteins that were substantially more visible in tris-NTA-DAP containing fractions than in control fractions after silver staining (A). Relative amount of protein in each band was quantified using ImageJ software. Association of the proteins at 60 kDa and 12 kDa are statistically greater for the tris-NTA-DAP-liposome than control liposomes (* p<0.05) (B). Competition to disassociate mKate from 1% tris-NTA-DAP, 5% mPEG liposomes by Hemopexin (HPX, light gray) or to associate mKate with liposomes after recovery from plasma (dark gray) (C). (HPX Competition n=2; mKate Addition n=3)