Quantitation of adsorbed plasma proteins on coated and non-coated PLGA (poly(lactic-co-glycolic acid)) nanoparticles by LC-MALDI

摘要

In this work we focused mainly on the analyses of the apolipoprotein family especially Apo E and Apo A1. Three to four proteins from this family could be identified from coated and uncoated nanoparticles e.g. ApoA1, ApoC4, ApoE and ApoH. For investigation and quantification by LC-MALDI two different LC column materials were compared: C-18 and Poros R2. For Poros R2 material a higher reproducibility could be observed. Moreover, the selectivity was in several cases higher for Poros R2. The reason for this may be the more effective removal of specific detergents. Also with regard to the quantification using the N-reactive iTRAQ-reagent Poros R2 was superior to C-18 material. Thus, it was possible to eliminate the commonly used SCX separation for desalting prior to reverse phase chromatography. Also for quantification, Poros R2 material showed no loss of selectivity. With both materials, C-18 and Poros R2, the previously reported composition of the corona [4] could be confirmed. The results from quantification of individual proteins on the nanoparticles are presented.