A Simple Method for Assessment of Human Anti-Neu5Gc Antibodies Applied to Kawasaki Disease
Figure 1
A simple method to detect overall human anti-Neu5Gc reactivity.
A. Human antibodies can bind to mouse glycoproteins from WT (Neu5Gc-positive) or Cmah-KO (Neu5Gc-deficient) mice revealing anti-Neu5Gc and/or general anti-mouse reactivity. B. Illustration of the principles of EIA method that can separate specific anti-Neu5Gc reactivity from anti-mouse reactivity. Human antibodies are pre-incubated with Cmah-KO sera to absorb out anti-mouse reactivity, and binding to WT sera then reveals Neu5Gc-reactivity (grey circles = mouse glycoproteins; purple diamonds = Neu5Ac-containing glycoconjugates; red diamonds = Neu5Gc-containing glycoconjugates). C. Treatment of WT sialo-glycoconjugates with mild periodate cleaves the sialic acid side chain. D. Periodate pre-treatment of glycocojugates can demonstrate sialic acid specific binding but has no effect on anti-mouse reactivity.