{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,27]],"date-time":"2025-10-27T12:33:14Z","timestamp":1761568394126,"version":"build-2065373602"},"reference-count":29,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2018,7,27]],"date-time":"2018-07-27T00:00:00Z","timestamp":1532649600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["P30\u2010ES002109","P30CCA14051"],"award-info":[{"award-number":["P30\u2010ES002109","P30CCA14051"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["aiche.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Biotechnology Progress"],"published-print":{"date-parts":[[2018,11]]},"abstract":"<jats:p>\n                    Gene\u2010specific promoter methylation is involved in gene silencing and is an important cancer biomarker. Cancer\u2010specific methylation patterns have been observed and clinically validated for numerous gene promoters, but the knowledge gleaned from this large body of work is currently under\u2010utilized in the clinic. Methylation\u2010specific PCR is currently the gold standard method for clinical methylation assessment, but several research groups have proposed hybridization\u2010based techniques which could be simpler to implement and provide more accurate results. However, the sensitivity of this easier alternative must be improved dramatically in order to compete with methylation\u2010specific PCR. Efficient sample capture is a key step in maximizing sensitivity, so here we investigate the key parameters involved in (i) maximizing the capture of gene\u2010specific target DNA molecules at the surfaces of functionalized, magnetic microparticles and (ii) recognizing DNA methylation using an engineered methyl\u2010CpG\u2010binding domain (MBD) protein. The magnetic bead density, the probe concentration, and the MBD concentration were very important for maximizing detection, and other variables such as the hybridization time also impacted the target capture efficiency but had a smaller effect on the overall methylation assay. The effect of genomic DNA on the capture of the target sequence was also investigated, and model methylated vs. unmethylated target sequences could be distinguished in the presence of 1 ng\/\u03bcL genomic DNA. The findings we report related to the underlying binding events involved in hybridization\u2010based epigenotyping can be leveraged in combination with the many signal amplification and detection approaches that are currently being developed. \u00a9 2018 American Institute of Chemical Engineers\n                    <jats:italic>Biotechnol. Prog<\/jats:italic>\n                    ., 34:1589\u20131595, 2018\n                  <\/jats:p>","DOI":"10.1002\/btpr.2644","type":"journal-article","created":{"date-parts":[[2018,4,25]],"date-time":"2018-04-25T06:25:04Z","timestamp":1524637504000},"page":"1589-1595","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["An examination of critical parameters in hybridization\u2010based epigenotyping using magnetic microparticles"],"prefix":"10.1002","volume":"34","author":[{"given":"Brooke E.","family":"Tam","sequence":"first","affiliation":[{"name":"Dept. of Chemical Engineering Massachusetts Institute of Technology, 77 Massachusetts Avenue Cambridge MA 02139"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yining","family":"Hao","sequence":"additional","affiliation":[{"name":"Dept. of Chemical Engineering Massachusetts Institute of Technology, 77 Massachusetts Avenue Cambridge MA 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