Glycan Profiling Analysis System using Lectin Microarrays
Transcript of Glycan Profiling Analysis System using Lectin Microarrays
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GlycoStationTM Reader 1200 LecChipTM
Glycan Profiling Analysis System using Lectin Microarrays
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Technology Platform:
GlycoStationTM
Lectin Microarray:
LecChipTM
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Glycan is a face of cells: Life doesn’t exist without glycans
Alberts et al. (eds.) Molecular biology of the cell (3rd ed.) p.502
P.M.Rudd et al., J. Mol. Biol. 293, 351-366
Membrane Proteins are heavily glycosylated
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The basic concept of GlycoStationTM is not full identification of glycan structures but identification of epitope images of glycans using a number of lectins specific to partial structure of glycans.
・・・・・Analyze signals generated from labeled glycoproteins hybridized to lectin microarrays (“LecChipTM”). The resulting fluorescence patterns are analyzed differentially with the GlycoStationTM Tools Pro.
Lectinn
Biological Inference of glycan structures using a set of lectins
Evanescent-field fluorescence-assisted lectin microarray: a new strategy for glycan profiling, A. Kuno, J. Hirabayashi et al., Nature Methods, vol2, No.11, November (2005) pp851-856
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Differentiation Factors
Glycan/Lectin: Kd 10-4〜10-7 M
Antigen/Antibody:10-6〜10-9 M
Avidin/Biotin: 10-12 M
Lectin
Glycan
Washing
Interaction between glycan and lectin is so weak Lose binding
Washing Free!!
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Differentiation Factors For any complex glycans (even for mixtures), GlycoStation is able to analyze those epitope images with high throughput, high sensitivity, and high accuracy without cleaving glycans from the carrier proteins.
Merits of GlycoStation ・No wash process(high repeatability) ・Real time liquid phase analysis ・Quantitave analysis without missing any weak interactions ・For both N- and O-glycans ・Identification of Isomers ・High Sensitivity(LOD) 100pg/mL Glycoprotein 100pM Glycan 500-1,000 Cell ・Applicable for crude samples ・High Versatility and Applicability N. Uchiyama et al., Proteomics 2008, 8, 3042-3050
Without wash With wash
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Major Applications
1. Monitoring of Biosimilar Drugs and
those Host Cells
2. Glyco-Biomarker Discovery and
Screening
3. Characterization of Stem and
Differentiated Cells
4. Intestinal Bacteria and Probiotics
5. Pathological Examination
What we can offer!
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Major Applications
Biosimilar Drugs
ーLecChip is a quickest wayー
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Major Applications Powerful enough to see differences in EPOs
EPO_ALPHAgfedcb EPO_BETAgfedcb EPO_NESPgfedcb
LTL 1
PSA 2
LCA 3
UEA_I
4
AO
L 5
AAL 6
MAL_I
7
SN
A 8
SSA 9
TJA
-I 1
0
PH
A(L
) 11
ECA 1
2
RCA120 1
3
PH
A(E
) 14
DSA 1
5
GSL-I
I 16
NPA 1
7
ConA 1
8
GN
A 1
9
HH
L 2
0
ACG
21
TxLC_I
22
BPL 2
3
TJA
-II
24
EEL 2
5
ABA 2
6
LEL 2
7
STL 2
8
UD
A 2
9
PW
M 3
0
Jacalin 3
1
PN
A 3
2
WFA 3
3
ACA 3
4
MPA 3
5
HPA 3
6
VVA 3
7
DBA 3
8
SBA 3
9
Cals
epa 4
0
PTL_I
41
MAH
42
WG
A 4
3
GSL_I_
A4 4
4
GSL_I_
B4 4
5
Norm
alize
d I
nte
nsi
ty (
Net
Inte
nsi
ty)
100
95
90
85
80
75
70
65
60
55
50
45
40
35
30
25
20
15
10
5
0
Tetra-antennary N-glycan
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Major Applications
Glyco-Biomarker
ーLecChip is a short-cutー
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Major Applications
Glycoprotein from Abnormal cells
Glycoprotein from Normal cells
Differentiation Disease, or development specific glycans
Core proteins are identical
Basic Concept to Investigate Glycan-related Bio-markers
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Major Applications Basic Glyco-BioMarker Development Pipeline
The most suspicious glycan
changes and key lectins
Lectin Array Analysis
SDS-PAGE
2D Electrophoresis LC-MS
Identification of
Carrier Proteins
Screening of 1st set of
Marker Candidates
High throughput Screening
(Antibody-overlay Lectin Microarray)
Large Scale Validation
Narrowing down
Marker Candidates
Searching papers
And Database Y Lectin-ELISA Assay
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Major Applications
Probiotics
ー Glycome of Intestinal Bacteriaー
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Major Applications
Lectin Microarray Reveaqls Binding Profiled of Lactobacillus casei Strains in a Comprehensive Analysis of Bacterial Cell Wall Polysaccharides, E. Yasuda et. Al. Applied and Environmental Microbiology, July 2011, p.4539-4546
Glycan Profiling of Lactobacillus
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Major Applications
Regenerative Medicine
ーLecChip classifies Cellsー
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Major Applications
Specific lectin biomarkers for isolation of human pluripotent stem cells identified through array-based glycome analysis, Yu-Chieh Wang, Shinya Yamanaka, Jeanne Loring et al., Cell Research advance online publication 6 September 2011; doi:10.1038/cr.2011.148(2011)
Pluripotent iPS, ES cells are clearly discriminated by LecChips
Differentiated cells
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Major Applications What medical treatments I should apply?
As a secondary criterion of pathological examination to judge cancer malignant grade, through differential glycan profiling of biopsy samples etc.
A reported example: uterine corpus cancer, G1, G2, and G3 G1=respond to anticancer drugs G3=not respond to anticancer drugs
Glycan profiling of endometrial cancers using lectin microarray, Y. Nishijima, M. Toyoda, M. Yamazaki-Inoue, T. Sugiyama, M. Miyazawa, T. Muramatsu, K. Nakamura, H. Narimatsu, A. Umezawa, and M. Mikami, Genes Cells, 2012 Oct. 17(10):826-36. doi: 10.1111/gtc. 12003
Drug efficacy changes
with malignancy
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Glycan Profiling technologies and
Services GlycoBiomarker Therapeutic Proteins
Stem Cells
Simple & Quick to use
Diagnosis
for your success!
Probiotics
http://www.glycotechnica.com/english/