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Articular chondrocytes from osteoarthritic knee joints of elderly, in vitro expanded in thermo-reversible gelation polymer (TGP), exhibiting higher UEA-1 expression in lectin microarray

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REGENERATIVE THERAPY 2020年 14卷 234-237页

作者： Katoh, Shojiro Fujimaru, Atsuki Senthilkumar, Rajappa Preethy, Senthilkumar Abraham, Samuel J. K. Edogawa Evolutionary Lab Sci Edogawa Ku Edogawa Hosp Campus2-24-18 Higashikoiwa Tokyo 1330052 Japan Edogawa Hosp Dept Orthopaed Surg Edogawa Ku 2-24-18 Higashikoiwa Tokyo 1330052 Japan Nichi In Ctr Regenerat Med NCRM Fujio Eiji Acad Terrain FEAT PB 1262 Chennai 600034 Tamil Nadu India JBM Inc Edogawa Ku 3-1-14 Higashikoiwa Tokyo 1330052 Japan Yamanashi Univ Fac Med 1110 Shimokato Chuo Yamanashi 4093898 Japan GN Corp Co Ltd 3-8 Wakamatsu Kofu Yamanashi 4000866 Japan

Autologous chondrocytes expanded, are used as tools of regenerative therapies for cartilage injuries. However, inability to maintain the hyaline phenotype both and post transplantation, remains one of the major hurdles for long term efficacy under clinical settings. We have reported earlier, hyaline phenotype maintenance of both human and rabbit chondrocytes for a long duration both when cultured conditions using a Thermo-reversible Gelation Polymer (TGP) scaffold-based methodology and post-transplantation animal model of cartilage damage. Having intrigued by such encouraging outcome, we in this study, analysed the similar TGP culture environment whether would be able to allow expansion of severe osteoarthritis affected cartilage tissue from elderly patients and evaluated the cells using lectin microarray characterization for pluripotency. Cartilage tissue were obtained from patients (n = 7; age: 60-85 years) undergoing total knee arthroplasty for severe osteoarthritis. Chondrocytes were isolated and cultured in two groups: i. conventional culture without scaffold (2D) and ii. using a TGP scaffold-based culture (3D) up to 18 weeks. In addition to earlier reported findings such as maintenance of hyaline phenotype having been confirmed in this study as well, surface glycoprotein analysis by lectin microarray demonstrated that the α1-2 Fuc recognition lectin (UEA-1) (marker reported in literature for pluripotent stem cells) was found to be more highly expressed in 3D culture compared to 2D culture and even increased over time in 3D culture. We have developed an environment where osteoarthritis affected chondrocytes from the elderly could be cultured up to 18 weeks using TGP scaffold which express pluripotent cell associated surface glycoproteins compared to the conventional methodology.

关键词： 2D, Two-dimensional 3D, Three-dimensional ACI, Autologous chondrocyte implantation CO2, Carbon dioxide Cartilage Chondrocytes ESC, Embryonic stem cells Hyaline phenotype lectin microarray MACI, matrix-associated chondrocyte implantation Osteoarthritis PBS, Phosphate-buffered saline Pluripotency TGP, Thermo-reversible gelation polymer Thermo-reversible gelation polymer (TGP) scaffold hPSCs, Human pluripotent stem cells iPSC, Induced pluripotent stem cells

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lectin microarray analyses reveal host cell-specific glycan profiles of the hemagglutinins of influenza A viruses

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VIROLOGY 2019年 527卷 132-140页

作者： Hiono, Takahiro Matsuda, Atsushi Wagatsuma, Takanori Okamatsu, Masatoshi Sakoda, Yoshihiro Kuno, Atsushi Natl Inst Adv Ind Sci & Technol Biotechnol Res Inst Drug Discovery Glycosci & Glycotechnol Res Grp Tsukuba Ibaraki 3058568 Japan Keio Univ Sch Med Dept Biochem Shinjuku Ku Tokyo 1608582 Japan Japan Bioind Assoc Project Utilizing Glycans Dev Innovat Drug Discov Chuo Ku Tokyo 1040032 Japan Hokkaido Univ Fac Vet Med Dept Dis Control Microbiol Lab Sapporo Hokkaido 0600818 Japan

Glycan structures on hemagglutinin (HA) of influenza A viruses have been analyzed previously to understand their significance. However, the formerly established methods using mass spectrometry present disadvantages such as procedure complexity, sensitivity, and throughput. Our study has established a novel method for analyzing glycan profiles of HA using lectin microarray techniques. We successfully obtained glycan profiles of HA starting from 1 ml of the 10(6) TCID50 samples through simple antigen enrichment using optimized immunoprecipitation. The profiles were reasonably consistent with known glycan structures of HA. Next, we compared glycan profiles of the HAs prepared from chicken embryos, MDCK, Vero, and A549 cells, and demonstrated the host cell-specific HA glycan profiles. Notably, the HA from MDCK cells was alpha 1-3 galactosylated. Our method provides a highly sensitive and simple procedure for glycan profiling of the viral glycoproteins, thereby paving way for direct glycan analyses of human- and animal-derived virions.

关键词： Influenza A virus lectin microarray Hemagglutinin Glycosylation alpha-Gal

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lectin microarray Combined with Mass Spectrometry Identifies Haptoglobin-Related Protein (HPR) as a Potential Serologic Biomarker for Separating Nonbacterial Pneumonia from Bacterial Pneumonia in Childhood

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PROTEOMICS CLINICAL APPLICATIONS 2018年第6期12卷 1800030-1800030页

作者： Yang, Lin Yang, Zhou Cheng, Lei Cheng, Juan Cheng, Li Sun, Yangyang Li, Wenting Song, Kai Huang, Weichun Yin, Yong Tao, Shengce Zhang, Qinghua Shanghai Childrens Med Ctr Dept Clin Lab Shanghai 200127 Peoples R China Wayen Biotechnol Shanghai Inc Shanghai 201203 Peoples R China Shanghai Jiao Tong Univ Shanghai Ctr Syst Biomed Key Lab Syst Biomed Minist Educ Shanghai 200240 Peoples R China Shanghai Childrens Med Ctr Dept Pulm Dis Shanghai 200127 Peoples R China Shanghai MOST Key Lab Hlth & Dis Genom Shanghai 201203 Peoples R China

Purpose Experimental design Etiological diagnosis of pediatric patients with community-acquired pneumonia is difficult. For therapy, one of the major problems is the difficulty in separating bacterial pneumonia which would benefit from antibiotics from nonbacterial pneumonia. Therefore, to identify potential biomarkers for distinguishing nonbacterial pneumonia from bacterial pneumonia are sought. lectin microarray containing 91 lectins is used to screen serums from pediatric patients with pneumonia. lectin-based pull-down assay combined with LC-MS/MS is used to identify the potential biomarkers. Results Conclusions and clinical relevance SNA-I, a lectin binding preferentially to alpha 2-6 linked sialic acid residues, shows higher binding signals (near 42 kDa) in the mycoplasma pneumonia group, when compared with the other groups. A total of 18 proteins are identified with LC-MS/MS. By western blot analysis, the authors confirm that the expression of haptoglobin-related protein (HPR) is elevated in pediatric patients with pneumonia compared with normal children (p < 0.001). Furthermore, HPR is higher in the mycoplasma pneumonia group (p < 0.01) and the viral pneumonia group (p < 0.05), when compared with the bacterial pneumonia group. These results indicate that HPR is a potential serologic biomarker which can differentiate between bacterial pneumonia and nonbacterial pneumonia. Detection of serum HPR might be useful for clinical diagnosis.

关键词： biomarker haptoglobin-related protein lectin microarray mass spectrometry pneumonia

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lectin microarray analysis of isolated polysaccharides from Sasa veitchii

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BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 2017年第9期81卷 1687-1689页

作者： Yagi, Hirokazu Tateno, Hiroaki Hayashi, Kyoko Hayashi, Toshimitsu Takahashi, Katsuhiko Hirabayashi, Jun Kato, Koichi Tsuboi, Masamichi Nagoya City Univ Grad Sch Pharmaceut Sci Nagoya Aichi Japan Natl Inst Adv Ind Sci & Technol Biotechnol Res Inst Drug Discovery BRD Tsukuba Ibaraki Japan Chubu Univ Inst Life & Hlth Sci Kasugai Aichi Japan Hoshi Univ Inst Med Chem Tokyo Japan Natl Inst Nat Sci Inst Mol Sci Okazaki Aichi Japan Natl Inst Nat Sci Okazaki Inst Integrat Biosci Okazaki Aichi Japan Univ Tokyo Grad Sch Pharmaceut Sci Tokyo Japan

We report lectin microarray profile of the polysaccharide fraction derived from Sasa veitchii leaf that exhibits anti-influenza activity. This fraction showed higher reactivities with lectins known as binders to oligo-mannose, fucose, or galactose. Our findings along with previously reported monosaccharide components suggest that the polysaccharide can be cross-reactive with cell surface receptors involved in immune system, thereby exerting anti-influenza activity.

关键词： lectin microarray polysaccharides Sasa veitchii anti-influenza virus activity

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The use of lectin microarray for assessing glycosylation of therapeutic proteins

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MABS 2016年第3期8卷 524-535页

作者： Zhang, Lei Luo, Shen Zhang, Baolin Food & Drug Adm Ctr Drug Evaluat & Res Off Biotechnol Prod Silver Spring MD USA

Glycans or carbohydrates attached to therapeutic glycoproteins can directly affect product quality, safety and efficacy, and therefore must be adequately analyzed and controlled throughout product life cycles. However, the complexity of protein glycosylation poses a daunting analytical challenge. In this study, we evaluated the utility of a lectin microarray for assessing protein glycans. Using commercial lectin chips, which contain 45 lectins toward distinct glycan structures, we were able to determine the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies. lectin binding signals were analyzed to generate glycan profiles that were generally consistent with the known glycan patterns for these glycoproteins. In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. These data suggest that lectin microarray could be used for screening glycan patterns of therapeutic glycoproteins.

关键词： monoclonal antibodies therapeutic glycoproteins lectin microarray Glycan analysis

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Fucosylation is associated with the malignant transformation of intraductal papillary mucinous neoplasms: a lectin microarray-based study

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SURGERY TODAY 2016年第10期46卷 1217-1223页

作者： Watanabe, Kiminori Ohta, Masayuki Yada, Kazuhiro Komori, Yoko Iwashita, Yukio Kashima, Kenji Inomata, Masafumi Oita Univ Fac Med Dept Gastroenterol & Pediat Surg Hasama Machi Oita 8795593 Japan Oita Prefectural Hosp Dept Pathol Oita Japan

Intraductal papillary mucinous neoplasm (IPMN) is an intraductal mucin-producing pancreatic neoplasm with the potential for malignant transformation. Changes in glycans expressed on the cell surface and glycotransferases play important roles in malignant transformation. We conducted this study to analyze glycan alterations in IPMNs by using a lectin microarray and to identify the factors associated with altered glycans and their relationships with malignant transformation. Using a lectin microarray, we evaluated glycan expression in 22 samples of IPMN with carcinoma, obtained from curative resections performed in our department. We also used immunohistochemistry to investigate fucosyltransferase 8 (Fut 8) protein expression, which is associated with glycan alterations in IPMNs. The lectin microarray demonstrated that only two lectins, Aleuria aurantia lectin (AAL) and Aspergillus oryzae l-fucose-specific lectin (AOL), which bind to fucose, exhibited significant sequential increases from normal pancreatic duct to adenoma and carcinoma. Similarly, Fut 8 protein expression, which is associated with AAL and AOL, sequentially and significantly increased from the normal pancreatic duct to adenoma and carcinoma. lectin microarray analysis suggested that fucosylation is associated with the malignant transformation of IPMNs.

关键词： IPMN Fucosylation lectin microarray AAL AOL

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Use of lectin microarray to differentiate gastric cancer from gastric ulcer

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World Journal of Gastroenterology 2014年第18期20卷 5474-5482页

作者： Wei-Li Huang Yang-Guang Li Yong-Chen Lv Xiao-Hui Guan Hui-Fan Ji Bao-Rong Chi Department of Hepatology the First Clinic Hospital of Jilin University Department of Gastroenterology Affiliated Hospital of Beihua University Laboratory of Medicinal Chemical Biology College of Life SciencesNankai University Department of General Surgery the Central Hospital of Jilin City

AIM: To investigate the feasibility of lectin microarray for differentiating gastric cancer from gastric ulcer. METHODS: Twenty cases of human gastric cancer tissue and 20 cases of human gastric ulcer tissue were collected and processed. Protein was extracted from the frozen tissues and stored. The lectins were dissolved in buffer, and the sugar-binding specificities of lectins and the layout of the lectin microarray were summarized. The median of the effective data points for each lectin was globally normalized to the sum of medians of all effective data points for each lectin in one block. Formalin-fixed paraffin-embedded gastric cancer tissues and their corresponding gastric ulcer tissues were subjected to Ag retrieval. Biotinylated lectin was used as the primary antibody and HRP-streptavidin as the secondary antibody. The glycopatterns of glycoprotein in gastric cancer and gastric ulcer specimens were determined by lectin microarray, and then validated by lectin histochemistry. Data are presented as mean +/- SD for the indicated number of independent experiments. RESULTS: The glycosylation level of gastric cancer was significantly higher than that in ulcer. In gastric cancer, most of the lectin binders showed positive signals and the intensity of the signals was stronger, whereas the opposite was the case for ulcers. Significant differences in the pathological score of the two lectins were apparent between ulcer and gastric cancer tissues using the same lectin. For MPL and VVA, all types of gastric cancer detected showed stronger staining and a higher positive rate in comparison with ulcer, especially in the case of signet ring cell carcinoma and intra-mucosal carcinoma. GalNAc bound to MPL showed a significant increase. A statistically significant association between MPL and gastric cancer was observed. As with MPL, there were significant differences in VVA staining between gastric cancer and ulcer. CONCLUSION: lectin microarray can differentiate the different

关键词： Gastric cancer Gastric ulcer lectin microarray lectin histochemistry Differentiate

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Discriminating Latent Autoimmune Diabetes in Adults from Type 2 Diabetes using Surface Plasmon Resonance Imaging in Combination with lectin microarray

Discriminating Latent Autoimmune Diabetes in Adults from Typ...

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The 6th International Conference on Nanoscience and Technology, China 2015 (第六届中国国际纳米科学技术会议)

作者： Wenfei Zhou Ling Zhu Mo Yang Jinsong Zhu National Center for Nanoscience and Technology Beijing 100190China University of the Chinese Academ National Center for Nanoscience and Technology Beijing 100190China

Diabetes mellitus is one of the most challenging health concerns in the *** at least 30%of the diabetic patients remaining undiagnosed or misdiagnosed,rapid and early diagnosis is of great concern [1].

关键词： lectin microarray Surface plasmon resonance imaging Type 2 diabetes Latent autoimmune diabetes in adults

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Comparison of orthogonal chromatographic and lectin-affinity microarray methods for glycan profiling of a therapeutic monoclonal antibody

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JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES 2015年 997卷 162-178页

作者： Cook, Matthew C. Kaldas, Sherif J. Muradia, Gauri Rosu-Myles, Michael Kunkel, Jeremy P. Hlth Canada Biol & Genet Therapies Directorate Ctr Biol Evaluat Ottawa ON K1A 0L2 Canada Univ Ottawa Dept Med Dept Biochem Microbiol & Immunol Ottawa ON Canada

The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. The monosaccharide contents of these glycans following acid hydrolysis were confirmed by HPAEC-PAD with monosaccharide standards. Glycosylation analysis by HILIC-FD after stoichiometric labelling with two different fluorescent tags (2-AA and 2-AB) enabled direct quantitation. The 2-AA- and 2-AB-labelled versions of the same glycan standard panel yielded distinctive separation profiles suitable for orthogonal identification of mAb glycans. Glycan profiling with the lectin microarray required partial denaturation of the intact mAbs to expose the sequestered Fc N-glycans. Glycosylation fingerprints were obtained using a fluorescently labelled antibody directed against human IgG Fc. Fluorescence intensities from the fingerprints were deconvoluted with a proprietary algorithm to obtain semi-quantitative "glycan structural class" information. Glycosylation analyses of the four mAb lots by these four methods, which separate and detect oligosaccharides according to different principles, provided complementary and corroboratory qualitative and quantitative information. The predominant N-linked structures were core-fucosylated asialo diantennary structures with varying galactosylation. There were also trace amounts of afucosyl and bisected glycans, but no detectable sialylation by any of the four methods. The therapeutic mAb demonstrated a high degree of consistency in the types and amounts of N-linked glycans in the four lots (<6% CV), and between all four analysis methods (<6

关键词： Monoclonal antibody Glycosylation Glycan Oligosaccharide HPAEC-PAD HILIC-FD lectin microarray Glycoscope

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Glycoprotein profiling of stem cells using lectin microarray based on surface plasmon resonance imaging

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ANALYTICAL BIOCHEMISTRY 2014年 465卷 114-120页

作者： Nand, Amita Singh, Vikramjeet Wang, Peizhe Na, Jie Zhu, Jingsong Natl Ctr Nanosci & Technol Beijing 100190 Peoples R China Univ Chinese Acad Sci Beijing 100049 Peoples R China Tsinghua Univ Sch Med Ctr Stem Cell Biol & Regenerat Med Beijing 100084 Peoples R China

lectin microarrays have emerged as a novel platform for glycan analysis during recent years. Here, we have combined surface plasmon resonance imaging (SPRi) with the lectin microarray for rapid and label-free profiling of stem cells. In this direction, 40 lectins from seven different glyco-binding motifs and three different cell lines-mouse embryonic stem cells (mESCs), mouse-induced pluripotent stem cells (miPSCs), and mouse embryonic fibroblast stem cells (MEFs)-were used. Pluripotent mouse stem cells were clearly distinguished from non-pluripotent stem cells. Eight lectins-DBA, MAL, PHA_E, PHA_L, EEL, AAL, PNA, and SNA-generated maximal value to define pluripotency of mouse stem cells in our experiments. The discriminant function based on lectin reactivities was highly accurate for the determination of stem cell pluripotency. These results suggested that glycomic analysis of stem cells leads to a novel comprehensive approach for quality control in cell-based therapy and regenerative medicine. (c) 2014 Elsevier Inc. All rights reserved.

关键词： lectin microarray SPRi Stem cell Glycoprotein Pluripotency

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