Figure 1.

(a) NDLP hydrodynamic radius was strongly dependent on the foundation ND-complex size 488 – AlexaFluor 488, 750 – XenoLights CF750, Epi – Epirubicin. (b) NDLP zeta-potential was nearly neutral for all complexes. (c) Confocal microscopy of NDLPs prior to sizing demonstrated strong colocalization of ND (AlexaFluor 555) and lipid (DID) fluorescent labels. (d) Flow cytometry shows the presence of a dual positive NDLP population in addition to free NDs (AlexaFluor 488 only) and empty liposomes (DID only) (quantified in Table 1). (e) Size exclusion chromatography, nearly all fractions obtained from a size exclusion column contained the fluorescence signature of both the NDs and lipid bilayer in varying ratios. (f) Indirect ELISA using the fluorescence signature of ND-AlexaFluo 488 confirmed successful antibody conjugation to NDLPs. (g) Protein gel electrophoresis of tangential flow filtrate showed bands at approximately160 kDa (black arrowhead), 100kDa and 55kDa. (h) Quantification of the 160 kDa band demonstrated loading of 7.74 μg antibody/mg ND. (i) Epirubicin loading into ND clusters was quantified using absorbance at 485 nm of the supernatant after centrifugation showed consistent drug loading of greater than 85% of the stock solution or 17.3% (wt).