Essentials in Glycan Profiling Analysis using Lectin Microarrays

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In the interpretation of glycan profiling patterns (i.e., glycan profiles) taken by lectin microarrays, I have summarized important things and procedures as follows.

1. Some sort of normalization is absolutely necessary in comparing glycan profiles differentially. One of the most useful normalization methods is "Average Normalization". In this case, all of the lectin signals are devided by the average of all lectins on the array, and for convenience, the values are then multiplied by 100.


2. And, the differences in glycan profiles are interpreted taking lectin binding characteristics and CV (coefficient of variation) into consideration. Usually the CV is less than 10% in a lot, and that of lot-to-lot variation gets a little bit bigger than this. Lectin binding specificity is not one-to-one relationship like an antigen-antibody reaction, but is fairly broader than that. So, we must be careful in the interpretation if other lectins with similar binding characteristics are reacting in the same way or not, taking each binding specificity into consideration. As for the lectin binding characteristics, please refer to Table 2 shown on page 4448 in the following paper "Lectin microarrays: concept, principle and applications" for your information. Lectin microarrays: concept, principle and applications


3. In some cases, enzymatic digestion is very useful in understanding glycan profiles. For instance, vague points in the interpretation get clearer by comparing before and after sialidase digestion and by repeating the enzymatic digestion with galactosidase, acetylhexosaminidase, mannosidase in sequence. A figure 4 shown on page 854 in the following paper "Evanescent-field fluorescence-assisted lectin microarray: a new strategy for glycan profiling" would be such a very good example. Evanescent-field fluorescence-assisted lectin microarray: a new strategy for glycan profiling



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このページは、Adminが2013年10月15日 11:59に書いたブログ記事です。

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