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Fig. 3.
Comparative analysis of Lectin Microarray and Lectin NanoProbeArray. The ASP3026 ASF and 6SF were labeled with HiLyte Fluor 555 dye and used for probing the lectin array either by flooding the array (Lectin Microarray) or piezoelectric liquid dispensing (Lectin NanoProbeArray). The reactivity of lectins (SNA, MAA, and DSA) on the array with a range of concentrations for ASF and 6SF were analyzed as described in Materials and methods. A sample image for one block on the array is shown with the graph. Each block consisted of 5 identical rows of 6 spots with alternate spots of lectins and PBS buffer (control). Error bars in the graph represent standard error of the mean (N = 5).
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Piezoelectric probing of Lectin NanoProbeArrays
The binding pattern of piezoelectric liquid dispensed ASF and 6SF during Lectin NanoProbeArray experiments correlated well the results from lectin blots and conventional array probing experiments (Fig. 2 and Fig. 3). ASF did not show any binding to the sialic acid recognizing lectins SNA and MAA, while the modified glycoform (6SF) showed specific binding to SNA but not MAA. Such subtle differences in sialylation status can have important biological implications; for example α(2-6) linked sialic acids on β1-integrin affects integrin-mediated signaling, modulates carcinoma differentiation [18] and has a role in the recognition of host cells by influenza virus [19].