Journal of capillary electrophoresis最新文献_第2页

A novel strategy to identify autoantigens by proteomic analysis of plasma IgG-bound proteins 通过血浆igg结合蛋白的蛋白质组学分析鉴定自身抗原的新策略

Journal of capillary electrophoresis Pub Date : 2019-01-01 DOI: 10.2198/JELECTROPH.63.15

Takuya Toki, Y. Kodera, Ryo Konno, Yoshiya Hirata, Tatsuya Saito, M. Shichiri

{"title":"A novel strategy to identify autoantigens by proteomic analysis of plasma IgG-bound proteins","authors":"Takuya Toki, Y. Kodera, Ryo Konno, Yoshiya Hirata, Tatsuya Saito, M. Shichiri","doi":"10.2198/JELECTROPH.63.15","DOIUrl":"https://doi.org/10.2198/JELECTROPH.63.15","url":null,"abstract":"Autoimmune mechanisms have been hypothesized to underlie a number of human disorders in which both disease pathogenesis and diagnostic biomarkers remain poorly understood. This is partly due to a lack of efficient techniques for identification of plasma autoantibodies associated with specific pathophysiological conditions. We have developed a novel proteomic methodology to comprehensively identify plasma IgG-bound proteins using liquid chromatography tandem mass spectrometry (LC-MS/MS) after denaturing enriched plasma IgG to solubilize and release low molecular weight proteins. In total, we identified 44 proteins using this method that were undetectable in unprocessed plasma, 21 of which were not identified in the Human Plasma Proteome Draft of 2017. Comparison of plasma IgGbound proteins between healthy subjects and patients with isolated adrenocorticotropic hormone deficiency, a rare endocrine disorder speculated to involve autoimmune mechanisms, revealed several distinct IgG-bound proteins specifically detected in patient plasma but not in healthy subjects. Our results suggest that solubilization of low molecular weight proteins bound to enriched plasma IgG and subsequent proteomic analysis by LC-MS/MS could provide a promising strategy for identification of autoantigens in human peripheral blood.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"8 Pt 2 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83974663","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Current status of cancer proteogenomics: a brief introduction 癌症蛋白质基因组学研究现状简介

Journal of capillary electrophoresis Pub Date : 2019-01-01 DOI: 10.2198/jelectroph.63.33

E. Hattori, T. Kondo

{"title":"Current status of cancer proteogenomics: a brief introduction","authors":"E. Hattori, T. Kondo","doi":"10.2198/jelectroph.63.33","DOIUrl":"https://doi.org/10.2198/jelectroph.63.33","url":null,"abstract":"Proteogenomics is a novel approach to understand the molecular backgrounds of diseases. In cancer research, proteomic studies have been conducted without using the genome data of individual samples. For example, a common public database has always been used to identify proteins by mass spectrometry. However, tumor genomes, even tumors of the same type of cancer, can differ considerably, and such differences affect the response to treatments. Thus, genomic backgrounds should be considered when identifying proteins by mass spectrometry. In cancer proteogenomics, a virtual proteome database is generated using the genome data of identical samples for the mass spectrometric identification of proteins reflecting genetic mutations, which are not common and not cited in the commonly used databases. Such proteins are candidate biomarkers and therapeutic targets. Although previous studies have reported software capable of translating genomic data to proteomic data, a standard protocol has not been established. In addition, the utility of proteogenomics has also not been established, and it is not self-evident that proteins with mutations unique to certain groups can be exploited for innovative treatments or to provide clues for the resolution of biological problems in cancers. Collaborative efforts by cancer researchers and specialists in mass spectrometry and bioinformatics are required for fruitful advancements.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90397253","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Identification of cantharidin as a drug candidate for glioblastoma by using a Connectivity Map–based approach 基于连接图的方法鉴定斑蝥素作为胶质母细胞瘤的候选药物

Journal of capillary electrophoresis Pub Date : 2019-01-01 DOI: 10.2198/JELECTROPH.63.9

Zhiwei Qiao, T. Kondo

引用次数: 1

Gel-based analysis of protein phosphorylation status by rapid fluorometric staining using TAMRA-labeled Phos-tag 使用tamra标记的phos标签进行快速荧光染色，凝胶基分析蛋白磷酸化状态

Journal of capillary electrophoresis Pub Date : 2019-01-01 DOI: 10.2198/JELECTROPH.63.25

Hiroshi Kusamoto, Emiko Kinoshita-Kikuta, Tomoyo Nishimura, T. Nagai, E. Kinoshita, T. Koike

{"title":"Gel-based analysis of protein phosphorylation status by rapid fluorometric staining using TAMRA-labeled Phos-tag","authors":"Hiroshi Kusamoto, Emiko Kinoshita-Kikuta, Tomoyo Nishimura, T. Nagai, E. Kinoshita, T. Koike","doi":"10.2198/JELECTROPH.63.25","DOIUrl":"https://doi.org/10.2198/JELECTROPH.63.25","url":null,"abstract":"SUMMARY Phosphorylation, one of the most common post-translational modifications of proteins, plays a critical role in many biological processes. We have previously developed several analytical methods for determining the phosphorylation status of certain proteins by using a phosphate-capturing binuclear metal complex known as Phos-tag. Here, we describe a novel method for the gel-based in vitro analysis of the phosphorylation status of a protein by a simple and rapid fluorometric staining method that uses a tetramethylrhodamine (TAMRA)-labeled Phos-tag derivative (TAMRA–Phos-tag). The entire staining protocol, which requires less than 2 h to complete, uses three buffer solutions for staining, washing, and dilution, respectively, at room temperature. The gel-based analysis of phosphoproteins in a polyacrylamide gel can be conducted by using a fluorescence imaging scanner with a 532-nm excitation laser and a 580-nm longpass emission filter. As a practical example of the use of the TAMRA–Phos-tag staining method, we examined the time course of dephosphorylation of ovalbumin by an alkaline phosphatase. In addition, inhibitor profiling of a tyrosine kinase Abl was performed by using an Abl-substrate (GST-Abltide) and an Abl-inhibitor (Imatinib).","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"12 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74092630","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 4

Screening of a growth inhibitor library of sarcoma cell lines to identify potent anti-cancer drugs 肉瘤细胞系生长抑制剂文库的筛选以鉴定有效的抗癌药物

Journal of capillary electrophoresis Pub Date : 2019-01-01 DOI: 10.2198/JELECTROPH.63.1

Zhiwei Qiao, T. Kondo

{"title":"Screening of a growth inhibitor library of sarcoma cell lines to identify potent anti-cancer drugs","authors":"Zhiwei Qiao, T. Kondo","doi":"10.2198/JELECTROPH.63.1","DOIUrl":"https://doi.org/10.2198/JELECTROPH.63.1","url":null,"abstract":"There is a need for novel drugs for sarcoma treatment. In the present study, to identify inhibitors with potential therapeutic utility in sarcomas, we screened the growth inhibitory effects of 361 inhibitors, including experimental reagents and anti-cancer drugs approved for use in non-sarcoma malignancies and those under clinical trials. The inhibitors were initially tested using 10 osteosarcoma cell lines. The half-maximal inhibitory concentration (IC50) of leptomycin B, actinomycin D, chetomin, and staurosporine was <100 nM in all the cell lines. As the promiscuous effects of staurosporine on kinases make it unsuitable for clinical applications, the other three inhibitors were tested in an additional 15 sarcoma cell lines derived from synovial sarcoma, fibrosarcoma, liposarcoma, rhabdomyosarcoma, malignant peripheral nerve sheath tumor, leiomyosarcoma, and Ewing’s sarcoma. The IC50 of leptomycin B and actinomycin D was <100 nM in all cell lines and that of chetomin was <100 nM in all but three synovial sarcoma cell lines. Although the clinical development of leptomycin B, a chromosomal region maintenance (CRM)1/exportin (XPO)1 inhibitor, was discontinued because of toxicity, a previous clinical trial revealed that other CRM1/XPO1 inhibitors, such as selinexor, have anti-tumor effects in sarcomas. Actinomycin D has proven clinical utility in the treatment of sarcomas. Chetomin disrupts the interaction of hypoxia-inducible factor-1 with the transcriptional coactivator p300 and its clinical utility has not been established in sarcomas. Chetomin exhibited growth inhibitory effects on sarcoma cells with different histological subtypes. Library screening is a powerful approach to detect the potential utility of anti-cancer drugs in sarcoma treatment.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"238 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77627883","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Urinary carboxylesterase 5A fragment as an early diagnostic marker of cat chronic kidney disease 尿羧酸酯酶5A片段作为猫慢性肾病的早期诊断标志物

Journal of capillary electrophoresis Pub Date : 2019-01-01 DOI: 10.2198/JELECTROPH.63.39

H. Maeda, Kana Kobayashi, Toshifumi Watanabe, M. Satoh, F. Nomura, K. Sogawa

{"title":"Urinary carboxylesterase 5A fragment as an early diagnostic marker of cat chronic kidney disease","authors":"H. Maeda, Kana Kobayashi, Toshifumi Watanabe, M. Satoh, F. Nomura, K. Sogawa","doi":"10.2198/JELECTROPH.63.39","DOIUrl":"https://doi.org/10.2198/JELECTROPH.63.39","url":null,"abstract":"SUMMARY Chronic kidney disease (CKD) is a common disorder and cause of death in cats. In the classification proposed by the International Renal Interest Society (IRIS), stage I and II CKD are difficult to diagnose accurately using markers, in comparison with normal controls. We recently described a simple and highly reproducible two-step method for identifying potential disease-marker candidates among low-abundance urine proteins. Urine samples were taken from 56 normal control cats as the control group and from 56 cats with CKD (stage I). A carboxylesterase 5A fragment and filaggrin-2 fragment were identified as two proteins with higher levels in normal control cats. The performance of the ELISA of urine carboxylesterase 5A fragment was satisfactory in terms of recovery (97.2–102.4%) and within-run (1.3–3.6%) and between-day (1.5–4.1%) reproducibility. Urine carboxylesterase 5A fragment levels were significantly greater in normal cats (3.4±0.6 mg/dL) than in CKD (stage I) (1.9±0.5 mg/dL) (p<0.001). A carboxylesterase 5A fragment may be useful as a complementary marker to P-Cre and BUN for detection of CKD (stage I).","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"42 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88461555","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 3

Meta-analysis of global gene-expression profiles identify molecular signatures for histological subtypes of sarcomas 全球基因表达谱的荟萃分析确定了肉瘤组织学亚型的分子特征

Journal of capillary electrophoresis Pub Date : 2018-01-01 DOI: 10.2198/JELECTROPH.62.21

Zhiwei Qiao, Cuneyd Parlayan, Shigeru Saito, T. Kondo

{"title":"Meta-analysis of global gene-expression profiles identify molecular signatures for histological subtypes of sarcomas","authors":"Zhiwei Qiao, Cuneyd Parlayan, Shigeru Saito, T. Kondo","doi":"10.2198/JELECTROPH.62.21","DOIUrl":"https://doi.org/10.2198/JELECTROPH.62.21","url":null,"abstract":"SUMMARY Sarcomas are rare mesenchymal malignancies and comprise over 50 histological subtypes. Sarcomas are not well studied because the number of cases of individual sarcoma is low. The utilization of public data, such as gene expression data, may allow for improvement in the novel discovery of sarcoma. In this study, to gain insight into histological subtypes of sarcoma from a public database, we performed a meta-analysis of the gene-expression profiles by survey-ing the data deposited in the Gene Expression Omnibus database from 2001 to 2014. The gene-expression data for 10 sarcoma subtypes and the gene-expression profiles for 1002 cases were selected for comparative analysis. Genes with histology-oriented molecular signatures were identified, and the results were verified by functional validation using gene oncology analysis. Pathway analysis suggested the existence of differential biological processes among sarcoma subtypes. Furthermore, as an application of the sarcoma gene expression datasets used in this study, we investigated the gene expression patterns of the targets of pazopanib to predict the response of sarcoma to pazopanib. We found that the gene expression distribution patterns of targets of pazopanib were without distinction among 10 subtypes of sarcoma. Taken together, we identified the tissue-specific genes of 10 subtypes of sarcoma by bioinformatics analysis; our results demonstrated the utility of sarcoma datasets in public databases and provide valuable information for future rare cancer research.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"53 1","pages":"21-29"},"PeriodicalIF":0.0,"publicationDate":"2018-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72949290","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Characterization of panel antibodies for classification of cancer type using novel antibody-based phosphoproteomics 基于新型抗体的磷酸化蛋白质组学用于癌症类型分类的抗体鉴定

Journal of capillary electrophoresis Pub Date : 2018-01-01 DOI: 10.2198/JELECTROPH.62.1

Tomohito Ayabe, Y. Motofuji, Asako Saito, Shinya Ayabe, M. Koike, Y. Kodera, T. Maeda, Y. Eishi, H. Komatsu

引用次数: 0

Protein fractionation for proteomics using the SAINOME-plate 使用SAINOME-plate进行蛋白质组学的蛋白质分离

Journal of capillary electrophoresis Pub Date : 2018-01-01 DOI: 10.2198/JELECTROPH.62.11

Yoko Ino, H. Kagawa, Tomoko Akiyama, Yusuke Nakai, Sakura Ito, M. Shimoda, Makiko Kawamura, H. Hirano, Y. Kimura

引用次数: 1

Identification of cephalomannine as a drug candidate for glioblastoma via high-throughput drug screening 通过高通量药物筛选鉴定脑啡胺作为胶质母细胞瘤的候选药物

Journal of capillary electrophoresis Pub Date : 2018-01-01 DOI: 10.2198/JELECTROPH.62.17

Zhiwei Qiao, T. Kondo

引用次数: 2