GlycoTechnica Ltd.

Applications

Application of GlycoStation™ Glycan Profiling Analysis Technology

The potential applications of glycan profiling analysis are numerous and wide ranging. GlycoTechnica currently sees that the following three areas are the highest priprity as short term applications of the technology:

A various types of samples can be applied onto LecChip™(GlycoTechnica's Lectin Microarrays). GlycoStation™ (Evanescent-field fluorescence excitation profiler) is a very powerful and versatile system designed for glycan profiling analysis to have a quickest, easiest and high sensitive analysis way.

The most important differentiating factor of GlycoStation is the sensitivity. As you know, LC-MS/MS is very powerful in determining carrier proteins and N-glycan structures, however, it is not strong enough to screen glyco-biomarkers from a small amount of serum sample because of the low sensitivity. Although GlycoStation can not identify glycan structures digitally, it can give you meaningful results from only a few milli-liter of serum sample for not only N-glycans but also O-glycans. That's a good example to explain you the merit of GlycoStation™. Also, we should emphaseize that lectin microarray is an easiest way to identify isomers, such as α2,3-Sia and α2,6-Sia, which is quite tough with a conventional LC-MS/MS system, though.

Outline of Application of GlycoStation™ and LecChip™link to PDF

Glycan Profling Analysis of Crude Samples

GlycoStation™ system is not just for purified samples, but also very effective for differential analysis of crude samples such as cell lysate, serum, and so forth. One of the good examples is a comparison between CHO and Lec1 mutant cells with the membrane faction. The difference in N-glycan related binders is relatively strong, whereas that of O-glycan related binders is relatively weak. The following is just some of the interesting information easily discovered through differential profiling. Signals from branched complex-type N-glycan binders (PHA(L), PHA(E), ACG), α2,3-Sialic acid binder(MAL I), lactose binder (RCA120) drastically decrease in Lec1 (shown in the graph of the lower left side). On the contrary, signals from high-mannose type N-glycan binders (GNA, HHL, PWM, Calsepa, PSA, LCA) increase in Lec1 (shown in the graph of lower right side). These observations are quite reasonable, taking into consideration of the lack of glycosyltransferase GlcNAc-T1 in Lec1 mutant cells.

Glycan Profiling Analysis of Crude Samples

Glycan Profling Analysis of EPOs

EPO(Erythropoietin) is a drug for amenia. It is well know that drug efficacy of biotechnology-based drugs such as EPO and IgG is greatly affected by the glycosylation. According to a paper published in Folio Pharmacol., Jpn, 131, pp192-199, the second generation EPO, of which glycosylation is modulated with introducing two more N-glycans, has higher homatopoietic effct comparing with the first generation one. The figure below shows compariosn of glycan profiles between the fisrt generation and the second generation EPOs (red color shows the first genetation and blue color shows the second generation). The difference shown by PHA-L suggests that tetra-antennary N-glycans are more expressed in the second generation, the strong signal of MAL_I indicates that those N-glycans are heavily terminated by &alpha2,3 Sia, the differences shown by AOL and AAL suggest that those N-glycans are more fucosylated in the second generation, and also you can find that there are some differences in polylactosamine binder LEL and lactosamin binder STL. Thus, we believe that lectin microarray is very powerful in comparing differences of glycosylation among biotechnology-based drugs and useful in accerelating drug R&D. Of course, it is also possible to do glycan profiling of secreted glycoproteins into culture supernatant.

Glycan Profiling Analysis of EPO

New Biomarker Development: Antibody Overlay Lectin Microarray Method

This is a quickest shortcut to investigate glycan structures of a target protein. That is because the fluorescence coming from the labeld antibody reflects interactions of glycans of the target protein with immobilized lectins directly. It is possible to study glycan structures of a specific protein from crude samples. For instance, Kato-et al. has actually investigated glycan structures of podoplanin which is a mucin-type sialoglycoprotein and acts as a platelet-aggregating factor. They used a highly ractive anti-podoplanin antibody: NZ-1 for this purpose. This method is a kind of sandwich assay. After the incubation of podoplanin sample on LecChip, biotinated NZ-1 was applied, and the fluorescence was measured by using a Cy3 labeled streptavidin. Please refer to a reference listed below.

Glycan Analysis using an Antibody Overlay Lectin Microarray Methods

Cell Surface Glycome Profling Analysis of Living Cells

It is well known that the cell surface glycome changes from species to species, with differentiation stages, and development of malignant variations. Our standard protocol is based on Cy3 labeling onto the protein part (actually to the amino-group) of glycoproteins. However, recently, Tateno et al. has succeeded in investigating cell surface glycome of living cells using GlycoStation. They labeled living cells metabolically by a Cell-Tracker Orange CMRA regent were applied directly onto LecChip. One of the biggest merits of this method is that it becomes possible to investigate not only glycoproteins but also glycolipids on cell surfaces. They applied this method to CHO and Lec mutant cells (Lec1, Lec2 and Lec.8), and demonstrated that the result coincides very well with the expectation inferred from the lack of specific glycosyltransferase.
They also applied the same method to K562 cells to see the difference of cell surface glycome before and after the differentiation, and demonstrated how powerful this technology is in terms of characterizing stem cells.

Glycan Profiling Analysis of Paraffin-embedded Tissue Arrays

Matsuda et al. have investigated glycan profiling of formalin-fixed parafin-embedded tissue samples. They succeeded in showing that good quality of glycan profiling is possible with a very small amount of tissue scratch section (1.5mm in diameter) which contains roughly only 500cells. So, this must be an ultra-sensitive method for glycan profiling. We believe that this protocol is useful to screen glycobiomerkers, because there are so many paraffin-embedded tissue samples stocked waiting for awaiking from deep sleep. Glycolipids are eliminated during the deparaffinizing process.

Glycan Profiling Analysis of Paraffin-embedded Tissue Arrays

Characterization of Stem Cells (hES, iPS, MSC etc.) and Regenerative Medicine

It was shown that mES(mouse ES), F9(mouse embryonic carcinoma cell), and differentiated cells by retinoic acid were clearly discriminated each other by appliying hierarchical clustering and principal component analyses to those glycan profiling patterns using LecChips. This result was presented at ISSCR 2009 by Dr. Toyoda and Dr. Umezawa, NCCHD, et al.

Stem Cell Characterization

Intestinal Bacteria and Probiotics

It is well know that intestinal bacteria are deeply involved in human immune activity, health, and longlife. It is also known that carbohydrate moiety in the cell wall of intestinal bacteria is playing a pivotal role as a possible immune modulator. Recently, it was demonstrated that lectin microarray is able to profile bacterial cell surface glycome quite simply and quickly by combining a labeling method using SYTOX Orange. This was a joint R&D result between Yakult Central Institute for Microbiological Research and AIST. This method is now attracting so much attention as a powerful method promoting probiotics related research and development.

References

  1. Major Depressive Disorder
    H. Yamagata, S. Uchida, K. Matsuo, K. Harada, A. Kobayashi, M. Nakashima, F. Higuchi, T. Watanuki, T. Matsubara, and Y. Watanabe, Altered plasma protein glycosylation in a mouse model of depression and in patients with major depression, J. of Affective Disorders, June 2018, Vol.233, pp-79-85, https://doi.org/10.1016/j.jad.2017.08.057.
  2. Exosomes Glycan Profiling Analysis
    A. Shimoda, Y. Tahara, S. Sawada, Y. Sasaki, and K. Akiyoshi, Glycan profiling analysis using evanescent-field fluorescence-assisted lectin array: Importance of sugar recognition for cellular uptake of exosomes from mesenchymal stem cells., BBRC, 2017 Sept. 23, 491(3)pp701-707. doi:10.1016/j.bbrc.2017.07126.
  3. Tissue Glycan Mapping
    X. Zou, M. Yoshida, C. Nagai-Okatani, J. Iwaki, B. Tan, K. Hagiwara, T.Sato, Y. Itakura, E.Noro, H. Kaji, M. Toyoda, Y. Zhang, H. Narimatsu, and A. Kuno, A stadardized method for lectin microarray-based tissue glycome mapping, Sci. Rep., 2017 Mar 6;7:43560. doi: 10.1038/srep43560.
  4. Colorectal cancer recurrence and ABA
    K. Nakajima, M. Inomata, H. Iha, T. Endoh, N. Shiraishi, K. Kashima, and S. Kitano, Establishment of new predictive markers for distant recurrence of colorectal cancer using lectin microarray analysis, Cancer Med., 2015 (Feb); 4(2): 293-302. doi: 10.1002/cam4.342.
  5. Gastric cancer recurrence and BPL
    T. Futsukaichi, T. Etoh, K. Nakajima, T. Daa, H. Shiroshita, N. Shiraishi, S. Kitano, and M. Inomata, Decreased expression of Bauhinia purpurea lectin is a predictor of gastric cancer recurrence, Surg Today, 2015 Oct; 45(10): 1299-306. doi: 10.1007/s00595-015-1127-1.
  6. Fucosylation and Intraductal papillary mucinous neoplasms
    K. Watanabe, M. Ohta, K. Yoda, Y. Komori, Y. Iwashita, K. Kashima, and M. Inomata, Fucosylation is associated with the malignant transformation of intraductal papillary mucinous neoplasms: a lectin microarray-based study, Surg Today, 2016 Oct; 46(10): 1217-23. doi: 10.1007/s00595-015-1299-8.
  7. Innate immune responses and Glycosylation
    M. Yamamoto-Hino, M. Muraoka, S. Kondo, R. Ueda, H. Okano, and S. Goto, Dynamic regulation of innate immune responses in Drosophia by Senju-mediated glycosylation, Proc. Natl Acad Sci USA, 2015 May 5; 112(18): 5809-14. doi:10.1073/pnas.1424514112.
  8. Biobetter Characterization
    M. Roucka, K.Zimmermann, M. Fido, and A. Nechansky, Application of Lectin Array Technology for Biobetter Characterization: Its Correlation with FcyRIII Binding and ADCC, Microarrays 2017, 6(1); doi:10.3390/microarrays6010001.
  9. Muchin-Like Glycopeptides
    G. Artigas, H. Hinou, F. G. Martin, H. J. Gabius, and S. Nishimura, Synthetic Mucin-Like Glycopeptides as Versatile Tools to Measure Effects of Glycan Structure/Density/Position on the Interaction with Adhesion/Growth-Regulatory Galectins in Arrays, Chem. An Asian Journal, DOI: 10:1002/asia.201601420.
  10. FDA Paper for The use of lectin microarray for assesing glycosylation of therapeutic proteins
    L. Zhang, S. Luo, and B. Zhang, mAbs (2016), DOI: 10.1080/19420862.2016.1149662.
  11. FDA Paper for Glycan analysis of therapeutic glycoproteins
    L. Zhang, S. Luo, and B. Zhang, mAbs (2015), DOI: 10.1080/19420862.2015.1117719.
  12. Cancer and O-Linked Glycosylation of Muc1
    A. Matsuda, A. Kuno, T. Nakagawa, Y. Ikehara, T. Irimura, M. Yamamoto, Y. Nakamura, E. Miyoshi, S. Nakamori, H. Nakanishi, C. Viwatthanasittiphong, P. Srivatanakul, M. Miwa, J. Shoda, and H. Narimatsu, Lectin Microarray-Based Sero-Biomarker Verfication Targeting Aberrant O-Linked Glycosylation on Mucin 1, Anal Chem, 2015 Jul 21; 87(14):7274-81. doi: 10.1021/acs.analchem.5b01329. Epub 2015 Jul 7.
  13. Lymph node metastasis of advanced gastric cacer
    K. Yamashita, A. Kuno, A. Matsuda, Y. Ikehara, N. Katada, J. Hirabayashi, H. Narimatsu, M. Watanabe, Lectin Microarray technology identifies specific lectins related to lymph node metastasis of advanced gastric cancer, Gastric Cancer, 2015 Apr.4, PMID: 25840959
  14. Development of lectin microarray
    J. Hirabayashi, Synthesiology - English edition Vol.7, No.2, p.105 (Sep. 2014)
  15. Profiling of Cell Surface Glycome of Fungi
    A. Shibazaki and T. Gonoi, Lectin microarray technique for glycomic profiling of fungal cell surfaces, Methods Mol Biol., 2014; 1200: 1200-94.
  16. Cancer Diagnosis
    H. Narimatsu, Synthesiology - English edition Vol.5, No.3, p.201 (Dec. 2012)
  17. Cancer-specific Glycosylated Podoplanin
    Y. Kato and M. K. Kaneko, Scientific reports 4, Article Number: 5924, doi:10.1038/srep05924
  18. Glycan Profiling of Choriocarcinoma
    Y. Kobayashi, K. Masuda, K. Banno, N. Kobayashi, K. Umene, Y. Nogami, K. Tsuji, A. Ueki, H. nomura, K. Sato, E. Tominaga, T. Shimizu, H. Saya, and D. Aoki, Glycan profiling of gestational choriocarcinoma using a lectin microarray.
  19. Novel Glycan Marker for Diabetic Nephropathy
    K. Inoue, J. Wada, J. Eguchi, A. Nakatsuka, S. Teshigawara, K. Murakami, D. Ogawa, T. Terami, A. Katayama, A. Tone, I. Iseda, K. Hida, M. Yamada, T. Ogawa, and H. Makino, Urinary Fetuin-A is a Novel Marker for Diabetic Nephropathy in Type 2 Diabetes Identifies by Lectin Microarray.
  20. Lectin Microarrays and glycodiagnostic agent
    A. Kuno, T. Sato, H. Shimazaki, S. Unno, K. Saitou, K. Kiyohara, M. Sogabe, C. Tsuruno, Y. Takehama, Y. Ikehara, and H. Narimatsu, Reconstruction of a robust glycodiagnostic agent supported by multiple lectin-assisted glycan profiling, Proteomics Clin Appl., 2013 May3, doi: 10.1002/prca.201300010.
  21. Cancer Marker for Cholangiocarcinoma
    A. Matsuda, A. Kuno, H. Matsuzaki, T. Kawamoto, T. Shikanai, Y. Nakanuma, M. Yamamoto, N. Ohkohchi, Y. Ikehara, J. Shoda, J. Hirabayashi, and H. Narimatsu, Glycoproteomics-based cancer marker discovery adopting dual enrichment with WFA for high specific glyco-diagnosis of cholangiocarcinoma, J. of Proteomics 85 (2013) p.1-11.
  22. Cancer Marker for Hepatocellular Carcinoma
    H. Kaji, M. Ocho, A. Togayachi, A. Kuno, M. Sogabe, T. Ohkura, H. Nozaki, T. Angata, Y. Chiba, H. Ozaki, J, Hirabayashi, Y. Tanaka, M. Mizokami, Y. Ikehara, and H. Narimatsu, J. of Proteome Res., 2013, 12 p.2630-2640.
  23. Glycan Profiling of Retinal Degradation
    Lectin Microarray Profiling and Relative Quantification of Glycome Associated With Proteins of Neonatal wt and rd1 Mice Retinae, S. Ahuja, Invest Ophathalmol Vis Sci. 2013 May 7; 54(5): 3272-80. doi: 10.1167/iovs.12-11363.
  24. Xenotransplant from pig to human
    A lectin array analysis for wild-type and α-Gal-knockout pig islets versus healthy human islets, S. Miyagawa, A. Maeda, S. Takeishi, T. Ueno, N. Usui, S. Matsumoto, T. Okitsu, M. Goto, and H. nagashima, Surg Today, 2013 Apr.3.
  25. Adult T-cell Leukemia
    Glycan Profiling of Adult T-Cell Leukemia (ATL) Cells with the High Resolution Lectin Microarrays, H. Iha and M. Yamada, "T-Cell Leukemia - Characteristics, Treatment and Prevention", book edited by Mariko Tomita, ISBN 978-953-51-0996-9, Published: February 20, 2013 under CC BY 3.0 license, DOI: 10.5772/55386.
  26. Lectin Microarrays
    Lectin Microarrays: concept, principle and applications, J. Hirabayashi, M. Yamada, A. Kuno and H. Tateno, Chemical Society Reviews, 2013, doi:10.1039/C3CS35419A.
  27. Liver Fibrosis and Glyco-biomarker
    A serum "sweet-doughnut" protein facilitaes fibrosis evaluation and therapy assessment in patients with viral hepatitis, A. Kuno, Y. Ikehara, Y. Tanaka, K. Ito, A Matsuda, S. Sekiya, S. Hige, M. Sakamoto, M. Kaga, M. Mizokami and H. Narimatsu, Scientific Reports 3, Article number: 1065, doi:10.1038/srep01065.
  28. Dynamic glycome shift on iPS
    Structural and quantitative evidence for dynamic glycome shift on production of induced pluripotent stem cells, K. Hasehira, H. Tateno, Y. Onuma, Y. Ito, M. Asashima, and J. Hirabayashi, Mol Cell Proteomics., 2012 Dec;11(12):1913-23. doi: 10.1074/mcp.M112.020586.
  29. Endometrial cancers
    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.
  30. Metastasis-associated changes in glycosylation
    Lectin array-based strategies for identifying metastasis-associated changes in glycosylation, S. Fry, B. Afrough, A. Leathem, and M. Dwek, Methods Mol Biol., 2012;878:276-72
  31. Barrett's esophagus
    Molecular imaging using fluorescent lectins permits rapid endoscopic identification of dysplasia in Barrett's esophagus, Bird-Lieberman EL, Neves AA, Lao-Sirieix P, O'Donovan M, Novelli M, Lovat LB, Eng WS, Mahal LK, Brindle KM, Fitzgerald RC., Nature Med. 2012 Jan 15. doi: 10.1038/nm.2616.
  32. Application to iPS
    Specific lectin biomarkers for isolation of human pluripotent stem cells identified through array-based glycomic analysis, YC. Wang, M. Nakagawa, I Garitaonandia, I. Slavin, G. Altun, RM Lacharite, KL Nozor, HT Tran, CL Lynch, TR Leonardo, Y. Lue, SE Peterson, LC Laurent, S. Yamanaka, and JF Loring, Cell Res. 2011 Nov;21(11):1551-63. doi: 10.1038/cr.2011.148. Epub 2011 Sep 6.
  33. A triplex lectin-santibody sandwich immunoassay for liver fibrosis:
    LecT-Hepa: A triplex lectin-antibody sandwich immunoassay for estimating the progression dynamics of liver fibrosis assisted by a bedside clinical chemistry analyzer and an automated pretreatment machine, A. Kuno, Y. Ikehara, Y. Tanaka, K.Saito, K. Ito, C. Tsuruno, S. Nagai, Y. Takahama, M. Mizokami, J. Hirabayashi, and H. Narimatsu, Clinica Chimica Acta 412 (2011) 1767-1772.
  34. Lectin Microarray Profiling of PSGL-1/mIgG:
    Pichia pastoris-produced mucin-type fusion proteins with multivalent O-glycan subsitution as targeting molecules for mannose-specific receptors of the immune system, A. Gustafsson, M. Sjoblom, L. Strindelius, T. Johansson, T. Fleckenstein, N. Chatzissavidou, L. Lindberg, J. Angstrom, U. Rova, and J. Holgersson, Glycobiology, 2011 Aug. 21(8), 1071-1086.
  35. Glycan Profiling of Intestinal Bacteria:
    Lectin microarray reveals binding profiles of Lactobacillus casei strains in a comprehensive analysis of bacterial cell wall polysaccharides, E. Yasuda, H. Tateno, J. Hirabayashi, I. Iino, and T.Sako, Appl. Eviron Microbiol., 2011 Jul. 77(13), 4539-4546.
  36. Lectin Microarray Profiling of Breast Cancer:
    Lectin Microarray Profiling of Metastatic Breast Cancers, S. A. Fry, B. Afrough, H. J. Lomax-Browne, J. F. Timms, L. S. Velentzis, A. J.C. Leathem, Glycobiology (2011) doi: 10.1093/glycob/cwr045, First published online: April 19, 2011.
  37. A review paper: A lectin-based practical approach to complex glycans:
    Lectin-based structural glycomics: A practical approach to complex glycans, J. Hirabayashi, A. Kuno, H. Tateno, Electrophoresis, 2011 May;32(10):1118-28. doi: 10.1002/elps.201000650.
  38. A possible biomarker for iNPH
    A unique N-glycan on human transferrin in CSF: a possible biomarker for iNPH, S. Futakawa, K. Nara, M. Miyajima, A. Kuno, H. Ito, H. Kaji, K. Shirotani, T. Honda, Y. Tohyama, K. Hoshi, Y. Hanzawa, S. Kitazume, R. Imamaki, K. Furukawa, K. Tasaki, H. Arai, T. Yuasa, M. Abe, H. Arai, H. Narimatsu, and Y. Hashimoto, Neurobiol Aging. 2011 Apr 1. [Epub ahead of print]
  39. Lectin Microarray analysis of MSC, ES, and iPS:
    Lectin microarray analysis of pluripotent and multipotent stem cells, M. Toyoda, M. Yamazaki-Inoue, Y. Itakura, A. Kuno, To. Ogawa, M. Yamada, H. Akutsu, Y. takahashi, S. Kanzai, H. Narimatsu, J. Hirabayashi, and A. Umezawa, Genes to Cells, Vol.16, Issue 1, 1-11, January (2011), DOI: 10.1111/j.1365-2443.2010.01459.x
  40. Stem Cell Characterization by the Cell Surface Glycome:
    Potential Linkages Between the Inner and Outer Cellular States of Human Induced Pluripotent Stem Cells, S. Saito, Y. Onuma, Y. Ito, H. Tateno, M. Toyoda, H. Akutsu, K. Nishino, E. Chikazawa, Y. Fukawatase, Y. Miyagawa, H. Okita, N. Kiyokawa, Y. Shimma, A. Umezawa, J. Hirabayashi, K. Horimoto, and M. Asashima, ISB2010, Suzhou, China, September 9-11, 2010, pp.381-388.
  41. New GlycoBiomarker for Cholangiocarcinoma:
    Wisteria floribunda Agglutinin-Positive Mucin 1 is a Sensitive Biliary Marker for Human Cholangiocarcinoma, A. Matsuda, A. Kuno, H. Matsuzaki, T. Irimura, Y. Ikehara, Y. Zen, Y. Nakamura, M. Yamamoto, N. Ohkuhchi, J. Shoda, J. Hirabayasi, and H. Narimatsu, Hepatology. 2010 Jul;52(1):174-82.
  42. Xenotransplantatio of GalT knockout pig:
    S. Miyagawa, S. Takeishi, A. Yamamoto, K. Ikeda, H. Matsunari, M. Yamada, M. Okabe, E. Miyoshi, M. Fukuzawa, and H. nagashima, Xenotransplantation, 2010 Jan. Vol 17(1), pp.61-70.
  43. Relationship between activities of IL23R-Fc protein and its glycan structure:
    Transient expression of an IL-23R extracellular domain Fc fusion protein in CHO vs. HEK cells results in improved plasma exposure, K. F. Suen, M. S. Turner, F. Gao, B. Liu, A. Althage, A. Slavin, W. Ou, E. Zuo, M. Eckart, T. Ogawa, M. Yamada, T. Tuntland, J. L. harris, and J. W. Trauger, Protein Expr. Purif. (2010), doi:10.1016/j.pep.2009.12.015.
  44. A Strategy for Discovery of Cancer Glyco-biomarkers:
    A Strategy for discovery of cancer glyco-biomarkers in serum using newly developed technologies for glycoproteomics, H. Narimatsu, H. Sawaki, A. Kuno, H. Kaji, H. Ito, and Y.Ikehara, FEBS Journal (2009), doi:10.1111/j.1742-4658.2009.07430.x
  45. Chemoenzymatic Synthesis of N-glycan Clusters and those Affinity to Lectin Microarrays:
    Chemoenzymatic Synthesis and Lectin Microarray Characterization of a Class of N-Glycan Clusters, W. Huang, D. Wang, M. Yamada, and Lai-Xi Wang, J. Am. Chem. Soc., Nov.16, 2009.
  46. GPIase Activity of tACE:
    Testicular Angiotensin-Converting Enzyme with Different Glycan Modification: Characterization on Glycosylphosphatidylinositol-Anchored Protein Releasing and Dipeptidase Activities, G. Kondoh, Ho. Watanabe, Y. tashima, Y. Maeda, and T. Kinoshita, J. Biochem. 2009 145(1)115-121, doi:10.1093/jb/mvn148.
  47. A Potential Marker for Hepatic progenitor cells:
    High levels of E4-PHA-reactive oligosaccharides: potential as marker for cells with characteristics of hepatic progenitor cells, N. Sasaki, K. Moriwaki, N.Uozomi, K. Noda, N. Taniguchi, A. Kameyama, H. Narimatsu, S. Takeishi, M. Yamada, N. Koyama, and E. Miyoshi, Glyconj J., DOI 10.1007/s10719-009-9240-2, May (2009).
  48. Strategy for Glycoproteomics:
    Strategy for Glycoproteomics: Identification of Glyco-Alteration Using Multiple Glycan Profiling Tools (dagger), H. Ito, A. Kuno, H. Sawaki, M. Sogabe, H. Ozaki, Y. Tanaka, M. Mizokami, JI. Shoda, T. Angata, T. Sato, J. Hirabayashi, Y. Ikehara, and H. Narimatsu., J Proteome Res., Mar 6;8(3), pp.1358-1367 (2009).
  49. Advanced Antibody-overlay Lectin Microarray Method:
    Focused differential glycan analysis with the platform antibody-assisted lectin profiling (ALP) for glycan-related biomarker verification. A. Kuno, Y. Kato, A. Matsuda, MK. Kaneko, H. Ito, K. Amano, Y. Chiba, H. Narimatsu, and J. Hirabayashi, Mol Cell Proteomics, Jan;8(1), p.99., 2009.
  50. Microarray Methods and Protocols: Chapter 9:
    Lectin Microarrays, Masao Yamada, "Microarray Methods and Protocols" edited by Robert S. Matson, CRC Press, Taylor &Francious Group, p.141 (2009).
  51. Optimization of High-sensitive Detection:
    Optimization of evanescent-field fluorescense-assisted lectin microarray for high-sensitive detection of monovalent oligozaccharides and glycoproteins, Noboru Uchiyama, Atsushi Kuno, Hiroaki Tateno, Yoshiko Kubo, Mamoru Mizuno, Midori Noguchi, and Jun Hirabayashi, Proteomics Vol.8, p.3042 (2008).
  52. Development of Data-mining System:
    Development of a data-mining system for differential profiling of cell glycoproteins based on lectin microarray, Atsushi Kuno, Yoko Itakura, Masashi Toyoda, Yoriko Takahashi, Masao Yamada, Akihiro Umezawa, and Jun Hirabayashi, Journal of Proteomics & Bioinformatics(JPB), Vol.1, p.68 (2008.5).
    Download: 834KBlink to PDF
  53. A Review Paper:
    Concept, Strategy and Realization of Lectin-based Glycan Profiling, J. Hirabayashi, J. Biochem. 144, pp.139-147 (2008).
  54. Glycan Profiling of Paraffin-embedded Tissue Arrays:
    Development of an all-in-one technology for glycan profiling targeting formalin-embedded tissue sections, Atsushi Matsuda, Atsushi Kuno, Hiroyasu Ishida, Toru Kawamoto, Jun-ichi Shoda and Jun Hirabayashi, Biochemical and Biophysical Research Communications,370, p.259 (2008).
  55. The Era of Glycan Profiling has come:
    Glycomics's Infinite Potential and Applications to Healthcare, M. Yamada, GOR, Vol.9, No.1, p.16 (2007).
    Download: 2.3MBlink to PDF
  56. Profiling of Living Cell Surface Glycome:
    A novel strategy for mammalian cell surface glycome profiling using lectin microarray, H. Tateno, T. Sato, H. Narimatsu, and J. Hirabayashi, Glycobiology, Vol.17, No.10, p.1138 (2007).
  57. Antibody-overlay Lectin Microarray Method:
    Inhibition of tumor cell-induced platelet aggregation using a novel anti-podaplanin antibody reacting with its platelet-aggregation-stimulating domain, Y. Kato, M. K. Kaneko, A. Kuno, N. Uchiyama, K. Amano, Y. Chiba, Y. Hasegawa, J. Hirabayashi, H. Narimatsu, K. Mishima, and M. Osawa, doi:10.1016/j.bbrc.2006.08.171.
  58. Application to Crude Samples:
    Application of Lectin Microarray to Crude Samples: Differential Glycan Profiling of Lec MutantsF Youji Ebe, Atsushi Kuno, Noboru Uchiyama, Shiori Koseki-Kuno, Masao, Takashi Sato, Hisashi Narimatsu and Jun Hirabayashi, J Biochem, 139(3), p.323 (2006).
  59. Frontal Affinity Chromatography:
    High-throughput analysis of lectin-oligosaccharide interactions by automated frontal affinity chromatography, Methods, S. Nakamura-Tsuruta, N. Uchiyama, and J. Hirabayashi, Enzymol., Vol.415, p.311 (2006).
  60. Principle of Glycan Profiling Technology and the Performance:
    Evanescent-field fluorescence-assisted lectin microarray: a new strategy for glycan profiling: Atsushi Kuno, Noboru Uchiyama, Shiori Koseki-Kuno1, Youji Ebe, Seigo Takashima, Masao Yamada & Jun Hirabayashi, Nature Methods Vol.2, No.11, p.851 (2005).

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