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The response to the high concentration (40 μg/μL) of BSA is probably ascribed to nonspecific interactions of F-lectins with a hydrophobic pocket of BSA. Such nonspecific interactions are not negligible especially for the case of WGA, but not for other lectins. Thus, these data suggested that the considerable positive response was an over 10% recovery ratio for lectins other than WGA and over 20% for WGA in the glycoprotein analysis.
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Serious variability of the recovery ratio between spots on the same plate was not observed (see Figure s-4). Similarly, it is shown that the response patterns were practically conserved between independent arrays (see Figure s-5).
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WGA is able to bind both the β-GlcNAc derivatives and the NeuNAc (sialic acid) derivatives as shown in Figure 1c. In contrast, GSL-II can bind only GlcNAc derivatives, but not NeuNAc derivatives. As shown in Figure 8, the mammalian cell lines responded at the WGA spots, but not at the GSL-II spots. Thus, it is reasonable to conclude that these cell lines include NeuNAc derivatives in greater amount than GlcNAc. In the matter of AAL, the great difference between the AAL response and the UEA-I response is probably due to the difference in the affinity of AAL and UEA-I for fucose. As shown in Figure s-2, AAL shows an affinity to fucose that is stronger by 10-fold than UEA-I. Based on these data, the amount of the fucosylated species may be roughly speculated.
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The detection sensitivity is adjustable by modulating the quencher concentration. As a typical example, Table s-1 shows that we can tune the detection sensitivity of glucose from 5 to 24 mM.
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The detection limit for a monosaccharide (glucose) was 1 mM; a glycoprotein (Ribo B) was 0.5 μg/μL (30 μM). The typical pattern for a bacteria (NM522) lysate was detected at 0.5 μg/μL (Figure s-6).
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These may not be so problematic, because the corresponding quenchers were easily synthesized by the reaction of 13 with various saccharides, which have a reducing terminal using the aminooxy-method (Scheme 1b), and the pair can be universally used for various analytes once it is optimized.
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