**2.9.5 Enzymatic deglycosylation**

HDL (500 µg) was lyophilized and dissolved in 100 µL 2-DE sample buffer (described earlier).The sample was then incubated with 20 U of PNGase F (Sigma-Aldrich, P7367) in 37 C over night. After incubation, sample were stored in -20ºC until 2-DE.

HDL was desalted in a PD-10 column and eluted in a 50 mM Na2HPO4, pH 6.0 buffer. A volume corresponding to 1200 µg of HDL proteins were then incubated with 10 U of Neuraminidase (Sigma-Aldrich, N3786) in 37 C for 4 hours. After incubation, sample was desalted in a PD-10 column with desalting buffer (NH4HCO3, 12mM, pH 7.1) and subsequently frozen in -70ºC. Sample was lyophilized before 2-DE.
