Figure 1. Generation of OVA-neo-glycoconjugates with LeX that confers binding of OVA to MGL1.
(A) A glycan profile of OVA was generated using a multidimensional normal phase nano-HPLC coupled with an electrospray ionization interface mass spectrometer with an intercalated nanofluorescence detector. The different glycan species, indicated by numbers, are shown on the right; their relative proportion is represented in a pie chart. (B) ELISA showing functional modification of OVA with LeX glycans, as detected with anti-LeX antibodies and resulting in binding of MGL1-Fc. Unconjugated OVA does not carry any ligands for MGL1. Modification of OVA with LeX did not alter the ability to bind to MR as illustrated by equal binding kinetics of MR-Fc to native OVA and OVA-LeX. OVA and OVA-LeX preparations contain similar amounts of OVA as detected with anti-OVA antibodies.
Figure 1—figure supplement 1. The MALDI-TOF/TOF mass spectrum of OVA-LeX (Red) shows an increase of 1,2 KDa compared to unconjugated OVA (Blue), corresponding to addition of two LeX molecules per OVA molecule.
