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We developed a new anion\u2010exchange column for high\u2010performance liquid chromatography (<jats:styled-content style=\"fixed-case\">HPLC<\/jats:styled-content>) with electrostatic and hydrophobic properties. Both cytosine and thymine, corresponding to methylated and unmethylated cytosine after bisulfite modification, respectively, are captured by electrostatic interaction and then discriminated from each other by their hydrophobic interactions. The <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> methylation levels of synthetic <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> were quantified accurately and reproducibly within 10 minutes without time\u2010consuming pretreatment of <jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content> products, and the measured values were unaffected by the distribution of methylated CpG within the synthetic <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> fragments. When the <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> methylation status of the <jats:italic><jats:styled-content style=\"fixed-case\">FAM<\/jats:styled-content>150A<\/jats:italic> gene, a marker of the CpG island methylator phenotype specific to clear cell renal cell carcinoma (cc<jats:styled-content style=\"fixed-case\">RCC<\/jats:styled-content>), was examined in 98 patients with cc<jats:styled-content style=\"fixed-case\">RCC<\/jats:styled-content>, bulk specimens of tumorous tissue including cancer cells showing <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> methylation of the <jats:italic><jats:styled-content style=\"fixed-case\">FAM<\/jats:styled-content>150A<\/jats:italic> gene were easily identifiable by simply viewing the differentiated chromatograms, even when the cancer cell content was low. Sixteen cc<jats:styled-content style=\"fixed-case\">RCC<\/jats:styled-content> showing <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> methylation more frequently exhibited clinicopathological parameters reflecting tumor aggressiveness (ie, a larger diameter, higher histological grade, vascular involvement, renal vein tumor thrombi, infiltrating growth, tumor necrosis, renal pelvis invasion and higher pathological <jats:styled-content style=\"fixed-case\">TNM<\/jats:styled-content> stage), and had significantly lower recurrence\u2010free and overall survival rates. These data indicate that <jats:styled-content style=\"fixed-case\">HPLC<\/jats:styled-content> analysis using this newly developed anion\u2010exchange column could be a powerful tool for <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> methylation diagnostics, including prognostication of patients with cancers, in a clinical setting.<\/jats:p>","DOI":"10.1111\/cas.13566","type":"journal-article","created":{"date-parts":[[2018,3,9]],"date-time":"2018-03-09T05:36:40Z","timestamp":1520573800000},"page":"1690-1700","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":24,"title":["Novel method for <scp>DNA<\/scp> methylation analysis using high\u2010performance liquid chromatography and its clinical application"],"prefix":"10.1111","volume":"109","author":[{"given":"Takuya","family":"Yotani","sequence":"first","affiliation":[{"name":"Department of Pathology Keio University School of Medicine Tokyo Japan"},{"name":"Tsukuba Research Institute Research and Development Division Sekisui Medical Co., Ltd. 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