glycan profiling analysis system using lectin microarrays

GlycoStationTM Reader 1200 LecChipTM

Glycan Profiling Analysis System using Lectin Microarrays

Technology Platform:

GlycoStationTM

Lectin Microarray:

LecChipTM

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

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

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!!

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

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!

Major Applications

Biosimilar Drugs

ーLecChip is a quickest wayー

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

Major Applications

Glyco-Biomarker

ーLecChip is a short-cutー

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

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

Major Applications

Probiotics

ー Glycome of Intestinal Bacteriaー

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

Major Applications

Regenerative Medicine

ーLecChip classifies Cellsー

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

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

Glycan Profiling technologies and

Services GlycoBiomarker Therapeutic Proteins

Stem Cells

Simple & Quick to use

Diagnosis

for your success!

Probiotics

http://www.glycotechnica.com/english/