{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T06:30:34Z","timestamp":1791268234735,"version":"4.1.0"},"reference-count":20,"publisher":"Wiley","issue":"13","license":[{"start":{"date-parts":[[2005,7,29]],"date-time":"2005-07-29T00:00:00Z","timestamp":1122595200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Proteomics"],"published-print":{"date-parts":[[2005,8]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The need to identify protein or peptide biomarkers <jats:italic>via<\/jats:italic> readily available biological samples like serum, plasma, or cerebrospinal fluid is often hindered by a few particular proteins present at relatively high concentrations. The ability to remove these proteins specifically, reproducibly, and with high selectivity is increasingly important in proteomic studies, and success in this procedure is leading to an ever\u2010increasing list of lower abundant proteins being identified in these biological fluids. The current work addresses some of the potential problems in depleting proteins in typical biomarker studies, including nonspecific binding during depletion procedures and whether low molecular weight (LMW) species bind to the column in a so\u2010called \u201csponge\u201d effect caused by the ability of albumin or other high\u2010abundant proteins to bind peptides or protein fragments. LC\u2010MS\/MS methods were applied to the comparative analysis of an IgG\u2010based immunodepletion method and a Cibacron blue (CB)\u2010dye\u2010based method, for specificity of removing targeted proteins (binding fraction), as well as for assessing efficiency of target removal. This analysis was extended to examine the effects of repeated use of materials (cycles of binding and elution), in order to assess potential for carryover of one sample to the next. Capacity studies and efficiency of protein removal from the serum samples were followed for the IgG\u2010based system using both immunochemical assays (ELISA) as well as LC\u2010MS\/MS methods. Additionally, the IgG\u2010based system was further characterized for the removal of LMW polypeptides by nonspecific binding. We conclude that the IgG\u2010based system provided effective removal of targeted proteins, with minimal carryover, high longevity, and minimal nonspecific binding. Significant differences are noted between the depletion techniques employed, and this should be considered based on the expectations set during experimental design.<\/jats:p>","DOI":"10.1002\/pmic.200402021","type":"journal-article","created":{"date-parts":[[2005,7,29]],"date-time":"2005-07-29T07:19:44Z","timestamp":1122621584000},"page":"3304-3313","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":246,"title":["Differences among techniques for high\u2010abundant protein depletion"],"prefix":"10.1002","volume":"5","author":[{"given":"Nina","family":"Zolotarjova","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James","family":"Martosella","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gordon","family":"Nicol","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jerome","family":"Bailey","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barry\u2005E.","family":"Boyes","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William\u2005C.","family":"Barrett","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2005,8,16]]},"reference":[{"key":"e_1_2_1_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibtech.2003.10.013"},{"key":"e_1_2_1_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibtech.2004.06.005"},{"key":"e_1_2_1_4_2","doi-asserted-by":"publisher","DOI":"10.1021\/pr049921p"},{"key":"e_1_2_1_5_2","doi-asserted-by":"publisher","DOI":"10.1002\/elps.200305909"},{"key":"e_1_2_1_6_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00118a027"},{"key":"e_1_2_1_7_2","doi-asserted-by":"publisher","DOI":"10.1002\/jmr.300030509"},{"key":"e_1_2_1_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1359-6446(03)02655-2"},{"key":"e_1_2_1_9_2","first-page":"184","volume":"15","author":"Hinerfeld D.","year":"2004","journal-title":"J. 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