Separation
of PA-labeled
glycans was carried out on a Shimadzu HPLC system equipped with a
fluorescence detector. In case of RP-HPLC, a Hypersil ODS column (C18;
Agilent) was used with 100 mM ammonium acetate, pH 4.0, and 30% (v/v)
methanol; a gradient of the latter (1% per minute) was programmed.
The column was calibrated daily in terms of glucose units (g.u.) with
a pyridylaminated partial dextran hydrolysate. For 2D-HPLC, selected
fractions were reapplied to a combined hydrophilic-interaction anionic-exchange
HPLC (HIAX, Dionex IonPac AS11) with an inverse gradient of acetonitrile
in 800 mM ammonium acetate, pH 3.85, as previously described.32 (link) For further details refer to theSupporting Information .
of PA-labeled
glycans was carried out on a Shimadzu HPLC system equipped with a
fluorescence detector. In case of RP-HPLC, a Hypersil ODS column (C18;
Agilent) was used with 100 mM ammonium acetate, pH 4.0, and 30% (v/v)
methanol; a gradient of the latter (1% per minute) was programmed.
The column was calibrated daily in terms of glucose units (g.u.) with
a pyridylaminated partial dextran hydrolysate. For 2D-HPLC, selected
fractions were reapplied to a combined hydrophilic-interaction anionic-exchange
HPLC (HIAX, Dionex IonPac AS11) with an inverse gradient of acetonitrile
in 800 mM ammonium acetate, pH 3.85, as previously described.32 (link) For further details refer to the
