Global N-glycan Site Occupancy of HIV-1 gp120 by Metabolic Engineering and High-Resolution Intact Mass Spectrometry
Glycan identification using native mass spectrometry
News from Dec 19, 2016
A vital step in HIV vaccine development strategies has been the observation that some infected individuals generate broadly neutralizing antibodies that target the glycans on the surface of HIV-1 gp120. These antibodies target glycan epitopes on viral envelope spikes and yet the positions and degree of occupancy of glycosyla-tion sites is diverse. Therefore, there is a need to understand glycosylation occupancy on recombinant immunogens. The sheer number of potential glycosylation sites and degree of chemical heterogeneity impedes assessing the global sequon occupancy of gp120 glycoforms.
For this publication we trapped the glycan processing of recombinant gp120 to generate homogenous glycoforms, to facilitate the occupancy assessment by intact mass spectrometry. We show that gp120 monomers of the BG505 strain contain either a fully occupied sequenz or miss one and sometimes two glycans across the molecule. This biosynthetic engineering approach enables the analysis of therapeutically important glycoproteins otherwise recalcitrant to analysis by native mass spectrometry.
go to article DOI: 10.1021/acschembio.6b00854