Chemistry Reference
In-Depth Information
Figure 14.6
A strategy for live cell glycome pro-
fi ling. Fluorescently labeled live cells can be
directly profi led by the scanner in the liquid
phase.
14.7
Conclusions
A central issue in the study of lectins has always been elucidation of their sugar-
binding specifi city, which is closely associated with their physiological functions.
In this sense, the history of lectin research can be regarded as that of advancement
in technologies for the investigation of lectin-carbohydrate interactions. Beginning
with the most commonly used method, the hemagglutination assay, various
methods have been adapted to glycotechnology. A robust technique is FAC, which
was fi rst introduced by Kasai as a quantitative chromatographic method for biomo-
lecular interactions. This simple technique is refi ned by strategic combination of
a HPLC technique with an FD system using PA- oligosaccharides (synthetic access
by strategies outlined in Chapter 3). As a result, lectin- carbohydrate interaction
analyses are feasible in a high-throughput manner. Thus, the concept of glycan
profi ling has become a reality by means of lectins, which had previously been
regarded as merely 'useful tools', as a supplement to glycan analysis (please see
also Chapter 25.2, especially Figure 25.2, for an example from tumor biology). As
tool in lectin microarrays for lectin-based glycomics they play important roles in
studies of glycoproteins and cells, and in biomarker discovery, with specifi city
analysis performed by glycan microarrays.
Summary Box
Conjugation of lectins to a resin does not only enable glycan purifi cation.
Frontal affi nity chromatography (FAC), together with calorimetry and other
binding assays, is a systematic means to investigate lectin specifi city and to
determine
K
d
values. FAC thus contributes to the construction of a fundamen-
tal lectin database - an essential prerequisite for the second stage (the lectin
microarray). The lectin microarray is an emerging technique for glycome analy-
sis with medical perspective. It requires no particular skill on the part of non-
specialized researchers.
