{"id":2375,"date":"2011-11-03T21:41:53","date_gmt":"2011-11-04T02:41:53","guid":{"rendered":"https:\/\/kermitmurray.com\/murray\/?p=2375"},"modified":"2024-02-20T09:39:00","modified_gmt":"2024-02-20T15:39:00","slug":"digestion-microfluidic-chip-for-maldi-tof-ms","status":"publish","type":"post","link":"https:\/\/kermitmurray.com\/research\/2011\/11\/digestion-microfluidic-chip-for-maldi-tof-ms\/","title":{"rendered":"Development of an Automated Digestion and Droplet Deposition Microfluidic Chip for MALDI-TOF MS"},"content":{"rendered":"\n
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J. Lee, H. Musyimi, S. Soper, K.K. Murray, “Development of an Automated Digestion and Droplet Deposition Microfluidic Chip for MALDI-TOF MS,” J. Am. Soc. Mass Spectrom.<\/em> 19<\/strong><\/em> (2008) 964\u2013972. doi:10.1016\/j.jasms.2008.03.015<\/a>.<\/p>\n\n\n\n

Abstract<\/h4>\n\n\n\n
\"Schematic<\/a>
Schematic of the PMMA microfluidic chip: (a) the bioreactor measured 40 mm \u00d7 100 \u00b5m \u00d7 100 \u00b5m and the solution of sample and matrix are loaded by syringe pumps. On the end of the bioreactor, coaxial tubes were sealed to mix digests with a matrix solution and to deposit onto a MALDI target plate (b) Assembled tryptic digest microfluidic chip: components including PMMA chip and cover slip, inlet and outlet connectors, capillary and stainless steel tube.<\/figcaption><\/figure><\/div>\n\n\n\n

An automated proteolytic digestion bioreactor and droplet deposition system was constructed with a plastic microfluidic device for off-line interfacing to matrix assisted laser desorption\/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The microfluidic chips were fabricated in poly(methyl methacrylate) (PMMA), using a micromilling machine and incorporated a bioreactor, which was 100 \u00b5m wide, 100 \u00b5m deep, and possessed a 4 cm effective channel length (400 nL volume). The chip was operated by pressure-driven flow and mounted on a robotic fraction collector system. The PMMA bioreactor contained surface immobilized trypsin, which was covalently attached to the UV-modified PMMA surface using coupling reagents N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC) and hydroxysulfosuccinimide (sulfo-NHS). The digested peptides were mixed with a MALDI matrix on-chip and deposited as discrete spots on MALDI targets. The bioreactor provided efficient digestion of a test protein, cytochrome c, at a flow rate of 1 \u00b5L\/min, producing a reaction time of ~24 s to give adequate sequence coverage for protein identification. Other proteins were also evaluated using this solid-phase bioreactor. The efficiency of digestion was evaluated by monitoring the sequence coverage, which was 64%, 35%, 58%, and 47% for cytochrome c, bovine serum albumin (BSA), myoglobin, and phosphorylase b, respectively.<\/p>\n<\/div><\/div>\n<\/div><\/div>\n","protected":false},"excerpt":{"rendered":"","protected":false},"author":1,"featured_media":729,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[134,85,5],"tags":[11,121,9,22],"class_list":["post-2375","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-lsu","category-microfluidic","category-publication","tag-asms","tag-digestion","tag-maldi","tag-microfluidics","entry"],"_links":{"self":[{"href":"https:\/\/kermitmurray.com\/research\/wp-json\/wp\/v2\/posts\/2375","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/kermitmurray.com\/research\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/kermitmurray.com\/research\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/kermitmurray.com\/research\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/kermitmurray.com\/research\/wp-json\/wp\/v2\/comments?post=2375"}],"version-history":[{"count":2,"href":"https:\/\/kermitmurray.com\/research\/wp-json\/wp\/v2\/posts\/2375\/revisions"}],"predecessor-version":[{"id":2377,"href":"https:\/\/kermitmurray.com\/research\/wp-json\/wp\/v2\/posts\/2375\/revisions\/2377"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/kermitmurray.com\/research\/wp-json\/wp\/v2\/media\/729"}],"wp:attachment":[{"href":"https:\/\/kermitmurray.com\/research\/wp-json\/wp\/v2\/media?parent=2375"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/kermitmurray.com\/research\/wp-json\/wp\/v2\/categories?post=2375"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/kermitmurray.com\/research\/wp-json\/wp\/v2\/tags?post=2375"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}