Combinatorial Chemistry & High Throughput Screening, 2011, 14, 711-719 711
1386-2073/11 $58.00+.00 © 2011 Bentham Science Publishers Ltd.
Lectin Microarrays: A Powerful Tool for Glycan-Based Biomarker
Shu-Min Zhou1,2, Li Cheng1,2, Shu-Juan Guo1,2, Heng Zhu*,3,4 and Sheng-Ce Tao*,1,2
1Shanghai Center for Systems Biomedicine, Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai
Jiao Tong University, Shanghai 200240, China
2State Key Laboratory of Oncogenes and Related Genes, Shanghai, 200240, China
3Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD
4The High Throughput Biology Center, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA
Abstract: Cell surfaces, especially mammalian cell surfaces, are heavily coated with complex poly- and oligosaccharides,
and these glycans have been implicated in many functions, such as cell-to-cell communication, host-pathogen interactions
and cell matrix interactions. Not surprisingly then, the aberrations of glycosylation are usually indicative of the onset of
specific diseases, such as cancer. Therefore, glycans are expected to serve as important biomarkers for disease diagnosis
and/or prognosis. Recent development of the lectin microarray technology has allowed researchers to profile the glycans
in complex biological samples in a high throughput fashion. This relatively new tool is highly suitable for both live cell
and cell lysate analyses and has the potential for rapid discovery of glycan-based biomarkers. In this review, we will focus
on the basic concepts and the latest advances of lectin microarray technology. We will also emphasize the application of
lectin microarrays for biomarker discovery, and then discuss the challenges faced by this technology and potential future
directions. Based on the tremendous progress already achieved, it seems apparent that lectin microarrays will soon
become an indispensible tool for glycosylation biomarker discovery.
Keywords: Glycans, glycomics, lectin microarray, live cell profiling, high throughput.
in eukaryotes are glycosylated . In particular, most of the
plasma membrane-localized and secreted proteins are
heavily glycosylated. The surfaces of most cells are thus
covered by a dense and diverse array of glycans, which
mediate a variety of biological processes, such as cell-to-cell
communication, cell-matrix interactions, host-pathogen
interactions, development, tumor invasion and metastasis.
Besides, about 2% of the human genome encodes proteins
that are in some way involved in glycan biosynthesis,
degradation, or transportation . A slight change in the
composition or structure of these glycans may thus lead to
dramatic changes in cell phenotype, and even cause disease,
such as the I-Cell disease  and Leukocyte-adhesion
deficiency type II (LAD II) .
It has been estimated that over 50% of proteins expressed
biological process, such as differentiation, development,
morphogenesis, and embryogenesis, may lead to drastic
shifts in glycan expression [5, 6]. As compared to proteins
from healthy tissues, proteins from disease tissues usually
show altered glycosylation [7, 8], which include a high-
degree of N-glycan branching, extended polylactosamine
In turn, even stable changes in the cells during diverse
*Address correspondence to these authors at the Department of
Pharmacology and Molecular Sciences, Johns Hopkins University School of
Medicine, Baltimore, MD 21205, USA; Tel: 410-502-0878; Fax: 410-502-
1872; E-mail: email@example.com or Shanghai Center for Systems
Biomedicine, Key Laboratory of Systems Biomedicine (Ministry of
Education), Shanghai Jiao Tong University, Shanghai 200240, China; Tel:
+86 21 34207069; Fax: +86 21 34207069;
chains, abnormal fucosylation, sialylation and sulfation.
These alterations could thus serve as biomarkers for disease
diagnosis [9, 10]. For example, AFP-L3, the fucosylated
variant of Alpha-fetoprotein (AFP), shows a much higher
specificity than AFP for identifying people who are a high
risk for hepatocellular carcinoma (HCC) .
There are a handful of tools and technologies that can be
used to study glycosylation, such as liquid chromatography
(LC) [12, 13], mass spectrometry (MS)  , capillary
electrophoresis (CE) [15, 16] and flow cytometry (FCM)
[17, 18]. These technologies can be used to identify protein
glycosylation sites, for deciphering complex glycan
structures, and even for profiling live cell surface glycans.
complicated sample preparation procedures, none of these
technologies are well-suited for high throughput and high
content glycosylation investigations.
glycosylation represents the
throughput technologies for better understanding the
underlying relationships of glycan aberrations and diseases
and for identification of novel biomarkers.
However, because these traditional approaches are time-
and labor-intensive, and often involve
The fact that
most complex protein
demands new high
glycans, and the parallel analysis capability of microarrays, a
novel high throughput technology - the lectin microarray -
has recently been developed [19-22]. Typically, a set of
lectins that recognize a variety of glycans are printed and
immobilized onto functionalized glass slides to form a lectin
microarray. Because of its miniaturized nature and capacity
Taking advantage of the specific binding of lectins to
712 Combinatorial Chemistry & High Throughput Screening, 2011, Vol. 14, No. 8 Zhou et al.
for high throughput analysis, a lectin microarray has
numerous advantages over the traditional technologies, as
demonstrated in a series of recent publications. For
examples, a number of lectin microarrays have been
successfully applied in a wide range of biological and
clinical studies, such as bacteria cell surface lipid
polysaccharide (LPS) profiling [20, 23, 24], clinical sample
analysis [25, 26], studies of host-pathogen interactions 
and mammalian cell surface glycan profiling .
potential for a wide range of applications, lecitn microarray
technology has already been reviewed by others [29-31].
Herein, we will only give a brief introduction to this
technology. We will focus more on the basic concepts and
discuss the latest advances. Specifically, we will emphasize
the applications of lectin microarrays for cell surface glycan
biomarker discovery and for analysis of clinical samples. We
will also discuss the challenges that lectin microarray
technology faces and possible future directions.
As a powerful high throughput technology with great
LECTIN MICROARRAY TECHNOLOGY
from plants and fungi  and the first evidence that lectins
could specifically bind to carbohydrates on mammalian cell
surfaces was provided in the 1950s . The number of
presently reported lectins is estimated to be approximately
1,000, and this number is still growing annually.
Lectins were first discovered more than 100 years ago
and the high throughput capability of microarrays, the first
lectin microarray carrying four lectins was introduced in
2005 . Similar to other types of microarrays, there are
four key steps for the fabrication and application of lectin
microarrays, namely (i) immobilization of lectins on the
functionalized microarray surfaces, (ii) labeling of the
sample, (iii) probing the cell or cell lysate on microarray
surface , and finally (iv) readout of results (Fig. 1).
By combining the specific binding of lectins to glycans
immobilizing a set of lectins onto a microscope slide using a
standard contact or non-contact microarrayer [34-36].
However, we and others observed that surface chemistry
plays a crucial role in the success of a particular assay. For
example, we showed that nitrocellulose-coated surfaces (e.g.,
FAST and PATH slides) are the best to carry out protein
acetylation reactions  and protein-DNA interactions ;
however, it produced too much background in a kinase assay
. Therefore, one has to first decide which slide surfaces,
such as aldehyde- and epoxy-derivatized glass surfaces,
hydrogel-coated slides, NHS-derivatized
nitrocellulose [41, 42], nickel-coated slides and streptavidin-
coated slides, would produce the best results in terms of
signal-to-background ratio (Fig. 1a). There are in fact two
common means of deposition that have been employed: (1)
lectins with amine groups are bound to the solid surface
through epoxy-functionalized or N-hydroxysuccinimidyl
(NHS)-derived esters, and (2) absorption and diffusion
within 3D nitrocellulose or hydrogel surfaces. However, the
lectins immobilized through the above two strategies are
Lectin microarrays are typically fabricated by
randomly oriented, and so a portion of the immobilized
lectins may lack the optimal orientation, native multimeric
quarternary structure, or optimal multivalent clustering of
carbohydrate recognition domains (CRD). To overcome the
disadvantages of random immobilization, glutathione-S-
transferase (GST) has been fused with recombinant lectins
. The purified lectins were printed and immobilized onto
glutathione (GSH) coated slide surfaces through the specific
and oriented binding of GST to GSH. This oriented
immobilization is expected to enhance the accessibility of
the lectins to the glycans, and thus improve the overall
performance of the lectin microarrays. Through molecular
cloning and recombination, researchers may can further
remove the crossreacting glycan portions of lectins while
leaving the CRD of lectins intact, and thereby greatly
improve the optimal orientation of lectins in their native
Sources of Lectins
fabrication at present because of their commercial
availability . The majority of lectins most frequently
employed are thus purified from plants, though lectins from
bacteria, fungi, or mollusca are also viable options (Fig. 1b).
However, application of these lectins suffers from inherent
glycosylation and batch-to-batch variation. To address these
problems, recombinant lectins, expressed in E. coli, and their
mutants have been actively developed. An additional benefit
of these recombinant lectins is that a GST-Hisx6 fusion tags
can be attached to the N terminus, and so the orientation of
lectins can be more easily controlled during the
immobilization process, which may further improve the
sensitivity . At the moment, there are very few vertebrate
lectins, let alone human lectins, that have been applied on
lectin microarrays, which may be a limiting factor in the
Plant lectins are an obvious choice for lectin microarray
Labeling and Readout
signal readout strategies in the DNA/oligo microarray fields
could be adopted for lectin microarrays. However, currently,
the dominant labeling strategy is still fluorescence [22, 46,
47] or its derivatives, such as evanescent-field fluorescence
[21, 26] and fluorescence resonance energy transfer (FRET)
 (Fig. 1d). Though fluorescence labeling is suitable for a
wide range of applications of lectin microarrays, several
disadvantages are need to be overcome, including the
potential variability in labeling efficiency among the
analytes. Besides, it has been observed that dead cells exhibit
stronger fluorescence after binding to lectins than live cells
. It has also been observed that pre-incubation of CD4+
human T-cells with a dye affected cell binding to
immobilized carbohydrates on solid surfaces . To
overcome these disadvantages, several other strategies have
been developed so as to work in a label-free mode, such as
scanning ellipsometry , surface plasmon resonance
(SPR) , and resonance light scattering . It is also
possible to use a simple optical microscope to evaluate the
number of cells bound to the lectin panel [22, 53].
Theoretically, all of the established sample labeling and
Lectin Microarrays Combinatorial Chemistry & High Throughput Screening, 2011, Vol. 14, No. 8 713
Lectin Microarrays for Glycosylation Studies
and highly efficient technology for determining the glycan
composition from a complex mixture. Using four lectins,
Angeloni et al. were first to show the feasibility of using
lectin microarrays to differentiate the terminal sugar epitopes
with good sensitivity . Currently, lectin microarray
technology has been applied to a wide range of studies,
including protein glycosylation analysis, live cell surface
glycan profiling, and clinical sample analysis for glycan
biomarker discovery (Fig. 2).
Lectin microarrays are a high throughput, easy-to-handle,
Live Cell Surface Glycan Profiling
in profiling the glycans of intact cells, including bacteria,
fungi [21, 23, 24], and mammalian cells. The dynamics of
cell surface glycosylation during growth, and after
As a technology, lectin microarrays have been validated
stimulation have also been monitored [19, 25, 53-55]. Hsu
and Mahal have designed a lectin microarray with 21 lectins
to recognize closely related E. coli strains, such as the
nonpathogenic strains JM101 and HB101 and the pathogenic
strains RS218, which was not possible using the traditional
assays (hemagglutination). This work revealed that
pathogenic bacterial strains had distinct binding signals and
different binding patterns from those of non-pathogenic
strains, which could provide a surface glycan fingerprint of
bacteria [23, 24].
However, some of the most exciting results are from the
studies of the mammalian cell surface glycans. To
differentiate cell types based on the accessible glyco-
epitopes on the cell surface, Zheng et al. compared the
binding affinities of mammalian cell lines BHK-21 and
Caco-2 using a lectin array with 6 lectins . These authors
found that BHK-21 cells bound to three out of six lectins
Fig. (1). The procedure for a typical lectin microarray experiment. (a) Lectins are delivered and immobilized on the microarray surfaces
through a variety of surface chemistries. (b) Lectins could be obtained from a wide range of sources. (c) The glycosylation status of a wide
range of biological samples are analyzed on lectin microarrays. (d) The results are recorded by either a standard fluorescence microarray
scanner or other labeling and recording methodologies.
Source of Lectin：
Labeling and Readout：
energy transfer (FRET)
714 Combinatorial Chemistry & High Throughput Screening, 2011, Vol. 14, No. 8 Zhou et al.
while Caco-2 cells bound to two additional lectins. It is
interesting to note that the comparisons were made between
a hamster cell line and a human cell line, and so their results
might reflect species-specific variations. In another study,
significant differences in carbohydrate expression were
observed on normal and tumorigenic human breast cell lines,
as well as on sublines differing in their tendency to “home”
to different tissues during metastasis . On the other hand,
Ebe and Tateno observed different glycan profiles between
normal Chinese hamster ovary cells and lymphatic
endothelial cells (LEC) with different glycosylation-
defective mutants [25, 55]. Following this clue, they found a
different profile of splenocytes prepared from wild type and
(1-3)-N-acetyl- ?-d-glucosaminyltransferase II knockout
mouse . Moreover, the authors also demonstrated that
the lectin microarrays are suitable for profiling dynamic
changes in the cell-surface glycome during chemically
induced differentiation of K562 cells.
Fig. (2). A variety types of samples could be analyzed by lectin
microarrays. These samples includeDifferent types of bacteria,
fungi, viruses, mammalian cells, proteins and lysates from either
cell cultures, animals or clinical specimens.
In an independent study, we have developed a high-
content lectin microarray containing 94 lectins, which is the
most comprehensive lectin microarray prepared to date. To
qualitatively profile the lectin binding specificity of 24
human cell lines, we developed a binary algorithm that
generates a “glycocode” for each cell lines. This has allowed
us to hierarchically cluster these cell lines on the basis of
their accessible glycan composition. Further analysis of the
cell surface glycan signatures resulted in predicting
mannose-dependent tropism of bacteria. This prediction was
further confirmed using experimental approach. Importantly,
we also successfully identified three new lectin biomarkers
(i.e., LEL, AAL and WGA) that differentiate breast cancer
stem-like cells from their parental cell line MCF7. To
demonstrate the usefulness of such a biomarker, we
employed a mouse model and showed that lectin-enriched
cancer stem cells from MCF7 population could indeed
produce bigger and more aggressive tumors in animals .
Clinically Related Study
technology is the analysis of clinical samples for biomarker
purposes. At the moment, there is a long list of glycoproteins
and glycans that have been reported as disease-related
biomarkers [7, 8, 56], which could be generally monitored
from the clinical samples. Because of its power for high
microarrays are well suited for disease related biomarker
To fully explore their power for biomarker discovery,
however, the current format of lectin microarrays will
require extensive improvement. Firstly, in many clinical
circumstances, due to the limited availability of the sample
quantity, the number of target cells or target proteins in the
sample is usually too low to be effectively detected by
ordinary lectin microarray procedures. Secondly, since the
binding affinity between lectins and glycans are usually low
(Kd is in the range of 10-3~10-6 M) , the washing step
may remove the analytes of interest from the immobilized
lectins on the microarray, especially when the analytes are
cells. In order to overcome these problems, researchers have
modified lectin microarray technology in many ways. For
one, antibody-assisted lectin profiling (ALP) has been
developed for detecting glycoproteins at very low
concentrations by lectin microarrays. Kuno et al. used this
method to analyze the glycan structures of the protein hPod,
which has been proposed to enhance the metastatic potential
of glioblastoma cells . Using ALP, this protein was
enriched by the aid of the appropriate antibody to
concentrate this target protein and made it possible to be
analyzed by lectin array.
An additional improvement is the evanescent-field
activated fluorescence detection system [21, 26], which is a
label-free detection platform. With this, interactions between
the analyte and lectins are monitored on the glass surface
immersed under buffer that enables real-time detection of
fluorescently labeled glycans. Since the evanescent field is
generated within 200 nm, the background level is extremely
low and washing steps are not necessary. It has been
reported that this detection system has by far the highest
sensitivity among the lectin microarray recording systems,
with reported limit of detection values of 100 pg/ml for
glycoprotein (asialofetuin) and 100 pM glycan (asialo-
biantennary N-glycan) probes, respectively . In a most
recent report, Kuno et al. further combined ALP and the
evanescent-field activated fluorescence detection system 
to detect the fibrosis-specific biomarker from a 0.5 μL of
sera from 125 patients. As a result, 12 selected lectins
reflected fibrosis-associated glyco-alteration of ?1-acid
glycoprotein (AGP), which is a specific glycoprotein
Another important application of lectin microarray
and fast analysis, lectin
Lectin Microarrays Combinatorial Chemistry & High Throughput Screening, 2011, Vol. 14, No. 8 715
associated with liver fibrosis. In addition, they also found
that the use of three lectins (i.e., AOL, MAL, and DSA) on
the array enhanced the diagnostic value for liver cirrhosis to
95% diagnostic sensitivity and 91% diagnostic specificity.
OVERVIEW AND OUTLOOK
glycosylation, the most abundant and complex form of
posttranslational modification, is a key regulator in the
functioning of lipids, proteins, and cells. As described above,
lectin microarray technology has been shown to be a suitable
tool for glycan profiling. The number of diverse protocols
and applications of functional lectin microarrays has grown
dramatically. The applications of lectin microarrays for
screening glycan biomarkers are summarized in Table 1.
There are approximately 100 lectins commercially available
with the ability to recognize a fraction of glycan structures
present either on the surface or inside microbial or
mammalian cells. Furthermore, there are several lectin
microarray platforms that have already been commercialized,
Advances in research over years have made it clear that
for example, Qproteome™ GlycoArray with 6 lectins
(QIAGEN, Hilden, Germany), GlycoImage® Lectin Array
with 12 lectins in a 96-well format (Galab, Geesthacht,
Germany), and GP Bio's LecChip™ with a panel of 45
lectins (Moritex, Mainz, Germany).
We have already witnessed the power of lectin
microarray technology for glycosylation related studies.
However, as to further explore its applicability for a wider
range of applications, we have to address several of its
current limitations. Firstly, it is necessary to expand the
current panel of lectins to vertebrate lectins, especially
human lectins. Most of the known and predicted human
lectins are ligands and receptors on plasma membranes or in
body fluids, which may function as regulators of cell
adhesion, as binders of soluble extracellular and intercellular
glycoproteins, as regulators of protein levels in the blood,
and as effectors of host–pathogen interactions. For example,
some lectins that are found on the surface of mammalian
liver cells are also believed to be responsible for the removal
of certain glycoproteins from the circulatory system [60, 61].
The Applications of Lectin Microarrays for Screening Glycan Biomarkers
Labeling and Readout Refs.
Comparing the specific binding of lectins of
two cell lines BHK-21 and Caco-2
Fluorescence and bright-field
Comparing the differences in carbohydrate
expression on normal and tumorigenic human
breast cell lines
Direct obsevation under a phase
contrast microscope without
Comparing the significant changes in binding
affinities with lectins of 5 microbial strains in
different culture media
Resonance light scattering
using gold nanoparticles
followed by silver deposition
Combining lectin microarray with LC-
MS/MS to discover the cell surface
glycoprotein markers of a glioblastoma-
derived stem-like cell lines
Fluorescently labelled cells
CFSE cell tracking dye.
Combining lectin microarray with LC-
MS/MS to investigateg the serum
glycoprotein profiling differences between
cirrhosis and early hepatocellular carcinoma.
Serum sample labeled with EZ-
link iodoacetyl-LC-biotin with
6 21 Bacteria
Comparing the different expression of
carbohydrates of the closely related E. coli
nonpathogenic strains and, pathogenic strains
Fluorescently labelled cells,
Comparing the different expression of
glycoproteins of CHO cells and their
glycosylation-defective Lec mutants
CMRA-labeled whole cells
Comparing the different expression of
glycoproteins of CHO cells and their
glycosylation-defective Lec mutants and K-
562 cells differentiated with sodium butyrate
CMRA-labeled whole cells
To comaparing the different levels of
sialylation of hPod protein from several cell
lines using antibody-assisted lectin
CMRA-labeled whole cells
To find the fibrosis-specific glyco-alteration
in the sera of 125 patients and to distinguish
the patients with cirrhosis from the patients
with chronic hepatitis.
CMRA-labeled whole cells
To discover the biomarkers for cancer stem-
Fluorescently labelled cells
CFSE cell tracking dye.
716 Combinatorial Chemistry & High Throughput Screening, 2011, Vol. 14, No. 8 Zhou et al.
Therefore, lectin microarrays with human lectins could
largely expand its applications to the studies of cell
signaling, differentiation, apoptosis, and the discovery of
disease biomarkers. A practical approach would be to clone
and purify all the known and predicated human lectins or
lectin-like proteins, the current number of which is ~100.
Thus, it would appear necessary to undertake large scale
explorations to discover new lectins and their biochemical
characterization, and collect as many lectins as possible to
enrich the lectin library.
Secondly, it is not feasible to obtain detailed information
of glycosylation patterns and investigate their functions in
very complex samples such as cell lysates or crude clinical
samples using only one of the current glycan profiling
approaches. Lectin microarrays, MS analyses, and HPLC all
provide unique abilities to identify alterations
glycosylation patterns. Hence, it seems reasonable to
develop integrated strategies combining several diverse
analytical technologies to determine glycan structures and
the glyco-alterations that occur on cell surfaces and/or on
proteins from cultures or patients. In fact, there have recently
been investigations that combined LC-MS/MS and lectin
microarrays to dissect glycoprotein markers [62, 63]. In one
study, two galactose-specific lectins (TKA and PNA) were
selected to capture the differentially expressed glycoproteins
from a stem cell-like glioblastoma neurosphere culture and a
traditional adherent glioblastoma cell line, and these were
then analyzed by LC-MS/MS (Fig. 3). As a result, 11 and 12
differentially expressed candidate glycoproteins were
captured and identified using TKA and PNA, respectively.
Liu et al.  used the same platform for a HCC study as to
identify glycoprotein biomarkers for early diagnosis. They
found that the signal intensities of AAL and LCA binding
were significantly higher in HCC, indicating an elevated
level of fucosylation. The authors further identified that C3,
CE, HRG, CD14 and HGF were possible biomarkers to
Fig. (3). The discovery of cell surface glycan related biomarkers and its related signaling pathway through the integration of lectin
microarrays and mass spectrometry. (a) Live normal cells and tumor cells are fluorescent labeled, (b) and compared on lectin microarrays.
(c) The lectins with specific cell binding activity are selected, (d) to purify the differentially expressed surface glycoproteins. (e) The purified
proteins are further identified by LC-MS. (f) Finally, the related signaling pathways of those proteins could be revealed, (g) and the
following functional study could be carried out.
Cell ACell B
Live cells probing
membrane protein membrane protein
Lectins selected with
Lectin Microarrays Combinatorial Chemistry & High Throughput Screening, 2011, Vol. 14, No. 8 717
distinguish early HCC from cirrhosis. Strategies based on
combining lectin microarrays and MS show tremendous
potential to meet the requirement of fast, low-cost, precise
and high throughput analysis and discovery.
Thirdly, perhaps the most exciting application of lectin
microarray technology is in the discovery of novel disease
biomarkers. However, the methods by which samples are
collected, managed, and shared (biobanking) from the clinic
lag far behind technology development. To assure the
success of the glycan biomarker discovery using lectin
microarrays, high quality and high content sample sets are a
prerequisite. There are already some organizations/consort-
iums specific for biobanking, such as BiobankUK and
Victorian Cancer Biobank. Nonetheless, although China has
the largest population and the richest clinical sample
resource, there is still no any internationally well-recognized
organization for clinical biobanking in china. To lead or at
least secure a prestigious position in the worldwide
competition of biomarker discovery in the future, and thus
make a more significant contribution to clinical research in
the long run, it is urgently required the Chinese researchers
make extensive efforts towards effective clinical biobanking.
In summary, with the knowledge that glycans play such a
crucial role in various biological processes and diseases,
understanding the role of glycan structure and function in
these fundamental processes is urgently required. Lectin
microarrays, as a fast, high throughput and affordable
technology for glycan profiling, appear well suited to make a
significant contribution to this end. It seems clear that lectin
microarrays will soon become an indispensible tool for
glycan biomarker discovery and other glycosylation related
Laboratory of Oncogenes and Related Genes (No. 92-10-12),
the State Key Development Program for Basic Research of
China (Grant No. 2010CB529205), the Program for New
Century Excellent Talents in University (Grant No. NCET-
09-551), the Shanghai “Phosphor” Science Foundation,
(Grant No. 10QA1403800), the National Natural Science
Foundation of China (Grant No. 31000388) and SRF for
ROCS, SEM to SCT, and the National Institute of Health
(RR020839, CA125807, and GM076102 to HZ).
This work is supported in part by grants of the State Key
LAD II = Leukocyte-adhesion deficiency type II
AFP = Alpha-fetoprotein
= Hepatocellular carcinoma
= Liquid chromatography
MS = Mass spectrometry
= Capillary electrophoresis
= Flow cytometry
LPS = Lipid polysaccharide
= Carbohydrate recognition domains
GST = Glutathione-S-transferase
GSH = Glutathione
= Fluorescence resonance energy transfer
= surface plasmon resonance
LEL = Lycopersicon Esculentum Lectin
= Aleuria Aurantia lectin
= Wheat germ agglutinin
ALP = Antibody-assisted lectin profiling
= Limit of detection
= Trichosanthes kirilowii agglutinin
PNA = Peanut agglutinin
= Lens culinaris agglutinin-A
= Aspergillus oryzae l-fucose-specific lectin
MAL = Maackia amurensis Lectin
= Datura stramonium agglutinin
= Lymphatic endothelial cell
C3 = Complement 3
= Histidine-rich glycorprotein
HGF = Hepatocyte growth factor
 Apweiler, R.; Hermjakob, H.; Sharon, N. On the frequency of
protein glycosylation, as deduced from analysis of the SWISS-
PROT database. Biochim. Biophys. Acta., 1999, 1473, 4-8.
Schachter, H.; Freeze, H.H. Glycosylation diseases: quo vadis?
Biochim. Biophys. Acta., 2009, 1792, 925-930.
Kollmann, K.; Pohl, S.; Marschner, K.; Encarnação, M.; Sakwa, I.;
Tiede, S.; Poorthuis, B.J.; Lübke, T.; Müller-Loennies, S.; Storch,
S.; Braulke, T. Mannose phosphorylation in health and disease.
Eur. J. Cell Biol., 2010, 89, 117-123.
Gazit, Y.; Mory, A.; Etzioni, A.; Frydman, M.; Scheuerman, O.;
Gershoni-Baruch, R.; Garty, B.Z. Leukocyte adhesion deficiency
type II: long-term follow-up and review of the literature. J. Clin.
Immunol., 2010, 30, 308-313.
Mahal, L.K. Glycomics: towards bioinformatic approaches to
understanding glycosylation. Anti.cancer Agents Med. Chem.,
2008, 8, 37-51.
Varki, A. Biological roles of oligosaccharides: all of the theories
are correct. Glycobiology, 1993, 3, 97-130.
Adham, S.A.; Coomber, B.L. Glucose is a key regulator of
VEGFR2/KDR in human epithelial ovarian carcinoma cells.
Biochem. Biophys. Res. Commun., 2009, 390, 130-135.
Guo, H.B.; Randolph, M.; Pierce, M. Inhibition of a specific N-
glycosylation activity results in attenuation of breast carcinoma cell
invasiveness-related phenotypes: inhibition of epidermal growth
factor-induced dephosphorylation of focal adhesion kinase. J. Biol.
Chem., 2007, 282, 22150-22162.
Haltiwanger, R.S.; Lowe, J.B. Role of glycosylation in
development. Annu. Rev. Biochem., 2004, 73, 491-537.
Van Dyken, S.J.; Green, R.S.; Marth, J.D. Structural and
mechanistic features of protein O glycosylation linked to CD8+ T-
cell apoptosis. Mol. Cell Biol., 2007, 27, 1096-1111.
Jawed Altaf Baig, J.A.; Junaid Mahmood Alam, J.M.; Syed Riaz
Mahmood, S.R.; Baig, M.; Shaheen, R.; Sultana, I.; Waheed, A..
Hepatocellular carcinoma (HCC) and diagnostic significance of A-
fetoprotein (AFP). J. Ayub. Med. Coll. Abbottabad, 2009, 21, 72-
718 Combinatorial Chemistry & High Throughput Screening, 2011, Vol. 14, No. 8 Zhou et al.
 Tomiya, N.; Awaya, J.; Kurono, M.; Endo, S.; Arata, Y.;
Takahashi, N. Analyses of N-linked oligosaccharides using a two-
dimensional mapping technique. Anal. Biochem., 1988, 171, 73-90.
Hase. S.; Ikenaka, T.; Matsushima, Y. Structure analyses of
oligosaccharides by tagging of the reducing end sugars with a
fluorescent compound. Biochem. Biophys. Res. Commun., 1978,
Kameyama, A.; Kikuchi, N.; Nakaya, S.; Ito, H.; Sato, T.;
Shikanai, T.; Takahashi, Y.; Takahashi, K.; Narimatsu, H. A
strategy for identification of oligosaccharide structures using
observational multistage mass spectral library. Anal. Chem., 2005,
Kamoda, S.; Nakanishi, Y.; Kinoshita, M.; Ishikawa, R.; Kakehi,
K. Analysis of glycoprotein-derived
glycoproteins detected on two-dimensional gel by capillary
electrophoresis using on-line concentration method. J. Chromatogr.
A, 2006, 1106, 67-74.
Kamoda, S.; Kakehi, K. Capillary electrophoresis for the analysis
of glycoprotein pharmaceuticals. Electrophoresis, 2006, 27, 2495-
Poulain, S.; Lepelley, P.; Cambier, N.; Cosson, A.; Fenaux, P.;
Wattel, E. Assessment of P-glycoprotein expression by
immunocytochemistry and flow cytometry using two different
monoclonal antibodies coupled with functional efflux analysis in
34 patients with acute myeloid leukemia. Adv. Exp. Med. Biol.,
1999, 457, 57-63.
Frojmovic, M.; Wong T. Dynamic measurements of the platelet
membrane glycoprotein IIb-IIIa receptor for fibrinogen by flow
cytometry. II. Platelet size-dependent subpopulations. Biophys. J.,
1991, 59, 828-837.
Pilobello KT, Krishnamoorthy L.; Slawek, D.; Mahal, L.K.
Development of a lectin microarray for the rapid analysis of protein
glycopatterns. Chembiochem, 2005, 6, 985-989.
Angeloni, S.; JRidet, J.L.; Kusy, N; Gao, H.; Crevoisier, F.;
Guinchard, S.; Kochhar, S.;
Glycoprofiling with micro-arrays of glycoconjugates and lectins.
Glycobiology, 2005, 15, 31-41.
Kuno, A.; Uchiyama, N.; Koseki-Kuno, S.; Ebe, Y.; Takashima, S.;
Yamada, M.; Hirabayashi, J. Evanescent-field fluorescence-
assisted lectin microarray: a new strategy for glycan profiling. Nat.
Methods, 2005, 2, 851-856.
Zheng, T.; Peelen, D.; Smith, L.M. Lectin arrays for profiling cell
surface carbohydrate expression. J. Am. Chem. Soc., 2005, 127,
Hsu, K.L.; Mahal, L.K. A lectin microarray approach for the rapid
analysis of bacterial glycans. Nat. Protoc., 2006, 1, 543-549.
Hsu, K.L.; Pilobello, K.T.; Mahal, L.K. Analyzing the dynamic
bacterial glycome with a lectin microarray approach. Nat. Chem.
Biol., 2006, 2, 153-157.
Ebe, Y.; Kuno, A.; Uchiyama, N.; Koseki-Kuno, S.; Yamada, M.;
Sato, T.; Narimatsu, H.; Hirabayashi, J. Application of lectin
microarray to crude samples: differential glycan profiling of lec
mutants. J. Biochem., 2006, 139, 323-327.
Uchiyama, N.; Kuno, A.; Koseki-Kuno, S.; Ebe, Y.; Horio, K.;
Yamada, M.; Hirabayashi, J. Development of a lectin microarray
based on an evanescent-field fluorescence principle. Methods
Enzymol., 2006, 415, 341-351.
Krishnamoorthy, L.; Bess, J.W.; Jr., Preston, A.B.; Nagashima, K.;
Mahal, L.K. HIV-1 and microvesicles from T cells share a common
glycome, arguing for a common origin. Nat. Chem. Biol., 2009, 5,
Tao, S.C.; Li, Y.; Zhou, J.; Qian, J.; Schnaar, R.L.; Zhang, Y.;
Goldstein, I.J.; Zhu, H.; Schneck, J.P. Lectin microarrays identify
cell-specific and functionally significant cell surface glycan
markers. Glycobiology, 2008, 18, 761-769.
Dai, Z.; Zhou, J.; Qiu, S.J.; Liu, Y.K.; Fan, J. Lectin-based
glycoproteomics to explore and analyze hepatocellular carcinoma-
related glycoprotein markers. Electrophoresis, 2009, 30, 2957-
Hirabayashi, J. Concept, strategy and realization of lectin-based
glycan profiling. J. Biochem., 2008, 144, 139-147.
Gupta G, Surolia A, Sampathkumar SG. Lectin microarrays for
glycomic analysis. OMICS, 2010, 14, 419-436.
Musshoff, F.; Madea, B. Ricin poisoning and forensic toxicology.
Drug Test Anal., 2009, 1, 184-191.
Sigrist, H.; Sprenger, N.
 Watkins, W.M.; Morgan, W.T. Specific inhibition studies relating
to the Lewis blood-group system. Nature, 1957, 180, 1038-1040.
Delehanty, J.B.; Ligler, F.S. Method for printing functional protein
microarrays. Biotechniques, 2003, 34, 380-385.
Delehanty, J.B. Printing functional protein microarrays using
piezoelectric capillaries. Methods Mol. Biol., 2004, 264, 135-143.
Jones, V.W.; Kenseth, J..R.; Porter, M.D.; Mosher, C.L.;
Henderson, E. Microminiaturized immunoassays using atomic
force microscopy and compositionally patterned antigen arrays.
Anal. Chem., 1998, 70, 1233-1241.
Lin, Y.; Lu, J.; Zhang, J.; Walter, W.; Dang, W.; Wan, J.; Tao,
S.C.; Qian, J.; Zhao, Y.; Boeke, J.D.; Berger, S.L.; Zhu, H. Protein
acetylation microarray reveals that NuA4 controls key metabolic
target regulating gluconeogenesis. Cell, 2009, 136, 1073-1084.
Shaohui Hu, S.; Xie, Z.; Onishi, A.; Yu, X.; Jiang, L.; Lin, J.; Rho,
H.; Woodard, C.; Wang, H.; Jeong, J.S.; Long, S.; He, X.; Wade,
H.; Blackshaw, S.; Qian, J.; Zhu, H. Profiling the human protein-
DNA interactome reveals ERK2 as a transcriptional repressor of
interferon signaling. Cell, 2009, 139, 610-622.
Zhu, J.; Liao, G.; Shan, L.; Zhang, J.; Chen, M.R.; Hayward, G.S.;
Hayward, D.; Desai, P.; Zhu, H. Protein array identification of
substrates of the Epstein-Barr virus protein kinase BGLF4. J.
Virol., 2009, 83, 5219-5231.
Kusnezow, W.; Jacob, A.; Walijew, A.; Diehl, F.; Hoheisel, J.D.
Antibody microarrays: an evaluation of production parameters.
Proteomics, 2003, 3, 254-264.
Kramer, A.B.; Roozendaal, C.; Dullaart, R.P. Familial occurrence
of subacute thyroiditis associated with human leukocyte antigen-
B35. Thyroid, 2004, 14, 544-547.
Stillman, B.A.; Tonkinson, J.L. FAST slides: a novel surface for
microarrays. Biotechniques, 2000, 29, 630-635.
Propheter, D.C.; Hsu, K.L.; Mahal, L.K. Fabrication of an oriented
lectin microarray. Chembiochem., 2010, 11, 1203-1207.
Rudiger, H.; Gabius, H.J. Plant lectins: occurrence, biochemistry,
functions and applications. Glycoconj. J., 2001, 18, 589-613.
Hsu, K.L.; Gildersleeve, J.C.; Mahal, L.K. A simple strategy for
the creation of a recombinant lectin microarray. Mol. Biosyst.,
2008, 4, 654-662.
Koshi, Y.; Nakata, E.; Yamane, H.; Hamachi, I. A fluorescent
lectin array using supramolecular hydrogel for simple detection and
pattern profiling for various glycoconjugates. J. Am. Chem. Soc.,
2006, 128, 10413-10422.
Patwa TH, Zhao J, Anderson MA, Simeone DM, Lubman DM.
Screening of glycosylation patterns in serum using natural
glycoprotein microarrays and multi-lectin fluorescence detection.
Anal. Chem., 2006, 78, 6411-6421.
Lee, M.R.; Park, S.; Shin, I. Protein microarrays to study
carbohydrate-recognition events. Bioorg. Med. Chem. Lett., 2006,
Nimrichter, L.; Gargir, A.; Gortler, M.; Altstock, R.T.; Shtevi, A.;
Weisshaus, O.; Fire, E.; Dotan, N.; Schnaar, R.L. Intact cell
adhesion to glycan microarrays. Glycobiology, 2004, 14, 197-203.
Carlsson J MM, Lundstrom I, Danielsson B, Winquist F.
Investigation of sera from various species by using lectin affinity
arrays and scanning ellipsometry. Anal. Chim. Acta., 2005, 530,
Mecklenburga, M.; Svitela, J.; Winquistb, F.; Gangb, J.; Ornsteinc,
K.; Deya, E.; Bina, X.; Hedborgb, E.; Norrbyc, R.; Arwinc, H.;
Lundströmb, I.; Danielsson, B. Differentiation of human serum
samples by surface plasmon resonance monitoring of the integral
glycoprotein interaction with a lectin panel. Anal. Chim. Acta.,
2002, 459, 25-31.
Gao, J.; Liu, D.; Wang, Z. Screening lectin-binding specificity of
bacterium by lectin microarray with gold nanoparticle probes. Anal.
Chem., 2010, 82, 9240-9247.
Chen, S.; Zheng, T.; Shortreed, M.R.; Alexander, C.; Smith, L.M.
Analysis of cell surface carbohydrate expression patterns in normal
and tumorigenic human breast cell lines using lectin arrays. Anal.
Chem., 2007, 79, 5698-5702.
Pilobello, K.T.; Mahal, L.K. Deciphering the glycocode: the
complexity and analytical challenge of glycomics. Curr. Opin.
Chem. Biol., 2007, 11, 300-305.
Tateno, H.; Uchiyama, N.; Kuno, A.; Togayachi, A.; Sato, T.;
Narimatsu, H.; Hirabayashi, J. A novel strategy for mammalian cell
surface glycome profiling using lectin microarray. Glycobiology,
2007, 17, 1138-1146.
Lectin Microarrays Combinatorial Chemistry & High Throughput Screening, 2011, Vol. 14, No. 8 719 Download full-text
 Zhang, X.; Wu, X.; Li, J.; Sun, Y.; Gao, P.; Zhang, C.; Zhang, H.;
Zhou, G. MDR1 (multidrug resistence 1) can regulate GCS
(glucosylceramide synthase) in breast cancer cells. J. Surg. Oncol.,
Bakry, N.; Kamata, Y.; Simpson, L.L. Lectins from Triticum
vulgaris and Limax flavus are universal antagonists of botulinum
neurotoxin and tetanus toxin. J. Pharmacol. Exp. Ther., 1991, 258,
Kuno, A.; Kato, Y.; Matsuda, A.; Kaneko, M.K.; Ito, H.; Amano,
K.; Chiba, Y.; Narimatsu, H.; Hirabayashi, J. Focused differential
glycan analysis with the platform antibody-assisted lectin profiling
for glycan-related biomarker verification. Mol. Cell Proteomics,
2009, 8, 99-108.
Atsushi Kuno, A.; Ikehara, Y.; Tanaka, Y.; Angata, T.; Unno, S.;
Sogabe, M.; Ozaki, H.; Ito, K.; Hirabayashi, J.; Mizokami, M.;
Narimatsu, H. Multilectin assay for detecting fibrosis-specific
glyco-alteration by means of lectin microarray. Clin. Chem., 2010.
 Ozaki, K.; Lee, R.T.; Lee, Y.C.; Kawasaki, T. The differences in
structural specificity for recognition and binding between
asialoglycoprotein receptors of liver and macrophages. Glycoconj.
J., 1995, 12, 268-274.
Banerjee, S.; Majumder, G.C. Homologous liver parenchymal cell-
cell adhesion mediated by an endogenous lectin and its receptor.
Cell Mol. Biol. Lett., 2010, 15, 356-364.
He, J.; Liu, Y.; Xie, X.; Zhu, T.; Soules, M.; DiMeco, F.; Vescovi,
A.L.; Fan, X.; Lubman, D.M. Identification of cell surface
glycoprotein markers for glioblastoma-derived stem-like cells using
a lectin microarray and LC-MS/MS approach. J. Proteome Res.,
2010, 9, 2565-2572.
Liu, Y.; He, J.; Li, C.; Benitez, R.; Fu, S.; Marrero, J.; Lubman,
D.M. Identification and confirmation of biomarkers using an
integrated platform for quantitative analysis of glycoproteins and
their glycosylations. J. Proteome Res., 2009, 9, 798-805.
Received: January, 27, 2011
Revised: March 21, 2011 Accepted: March 27, 2011