A Simple Method for Assessment of Human Anti-Neu5Gc Antibodies Applied to Kawasaki Disease
Figure 2
Sialic acid characteristics of sera from WT and Cmah-KO Mice. A.
DMB-HPLC analysis of mouse sera confirms that WT is Neu5Gc-positive while Cmah-KO is Neu5Gc-deficient. B. Lectin binding on mouse sera demonstrates both Siaα2-6 (SNA) and Siaα2-3 (MAL1/2) glycoconjugates in both WT and Cmah-KO. After periodate treatment the signal is reduced demonstrating Sia specificity. The periodate-resistant signal suggest 9-O-acetylated Sias. C. Pre-treatment of coated-wells with base removes periodate resistant signals (white bars in B compared to C), confirming that some of the Sia are modified with 9-O-acetylation. D. Mass-spectrometry analysis of N-linked and O-linked glycans of WT and Cmah-KO sera reveals the most common sialoglycan structures. Each figure is representative of at least two independent experiments. Bars are mean ± s.d. of triplicates (A–C).