{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T22:21:36Z","timestamp":1778710896597,"version":"3.51.4"},"reference-count":26,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2006,6,30]],"date-time":"2006-06-30T00:00:00Z","timestamp":1151625600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotech &amp; Bioengineering"],"published-print":{"date-parts":[[2006,12,20]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Tethering of growth factors to biomaterial substrates via a polyethylene glycol (PEG) spacer has been established as a means of controlling dosage and conformation of the protein at the material surface, while retaining biological activity. However, the extent of modification through a comparison of bound versus unbound protein has not generally been characterized. In this work, covalent tethering of epidermal growth factor (EGF) to allylamine plasma modified polydimethylsiloxane (PDMS) substrates is characterized to determine the nature of the bound growth factor and to optimize the conditions for the reaction. Tethering is achieved via conjugation of EGF with homobifunctional <jats:italic>N<\/jats:italic>\u2010hydroxysuccinimide (NHS) ester of PEG\u2010butanoic acid (SBA<jats:sub>2<\/jats:sub>\u2010PEG) in solution, followed by exposure of the pegylated EGF to the aminated surfaces (solution first reaction). SDS\u2013PAGE analysis indicates that a low ratio of EGF:PEG is required to maximize the yield of the EGF\u2013PEG reaction; a relatively short reaction time is needed to limit hydrolysis of the NHS ester. With increasing amounts of PEG and a higher reaction time, a higher fraction of the EGF can be covalently tethered to the surfaces, as shown by binding of <jats:sup>125<\/jats:sup>I\u2010labeled EGF and subsequent washing with sodium dodecyl sulfate (SDS) to remove adsorbed protein. However, even under the optimal reaction conditions established by the SDS\u2013PAGE analysis, higher molecular weight EGF\u2013PEG complexes are observed by SDS\u2013PAGE and matrix\u2010assisted laser desorption\/ionization (MALDI). The presence of these complexes, as well as unreacted growth factor, can lead to a surface of heterogeneous composition. While these surfaces were found to have biological activity, stimulating the adhesion and growth of corneal epithelial cells versus PDMS controls, further optimization of reaction conditions, including the use of a homobifunctional PEG linker and possibly separation of reaction species are required to achieve a uniformly active and well\u2010defined biomaterial surface. \u00a9 2006 Wiley Periodicals, Inc.<\/jats:p>","DOI":"10.1002\/bit.21083","type":"journal-article","created":{"date-parts":[[2006,6,30]],"date-time":"2006-06-30T15:09:03Z","timestamp":1151680143000},"page":"1158-1166","source":"Crossref","is-referenced-by-count":28,"title":["Characterization of EGF coupling to aminated silicone rubber surfaces"],"prefix":"10.1002","volume":"95","author":[{"given":"Bettina J.","family":"Klenkler","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Heather","family":"Sheardown","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2006,6,30]]},"reference":[{"key":"e_1_2_1_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0928-4931(00)00157-0"},{"key":"e_1_2_1_3_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1007502828372"},{"key":"e_1_2_1_4_1","doi-asserted-by":"publisher","DOI":"10.1021\/bc990001k"},{"key":"e_1_2_1_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-4636(19980315)39:4<539::AID-JBM6>3.0.CO;2-K"},{"key":"e_1_2_1_6_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-4636(19981215)42:4<597::AID-JBM16>3.0.CO;2-R"},{"key":"e_1_2_1_7_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1025034925152"},{"key":"e_1_2_1_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0378-5173(02)00636-1"},{"key":"e_1_2_1_9_1","doi-asserted-by":"publisher","DOI":"10.1021\/la020027r"},{"key":"e_1_2_1_10_1","doi-asserted-by":"publisher","DOI":"10.1021\/bp0341390"},{"key":"e_1_2_1_11_1","doi-asserted-by":"publisher","DOI":"10.1002\/jbm.820251211"},{"key":"e_1_2_1_12_1","doi-asserted-by":"publisher","DOI":"10.1021\/la981356f"},{"key":"e_1_2_1_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0928-4931(98)00061-7"},{"key":"e_1_2_1_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-4827(02)00098-8"},{"key":"e_1_2_1_15_1","doi-asserted-by":"publisher","DOI":"10.1002\/jbm.a.10168"},{"key":"e_1_2_1_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/0169-409X(93)90005-O"},{"key":"e_1_2_1_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0142-9612(01)00334-9"},{"key":"e_1_2_1_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biomaterials.2005.05.045"},{"key":"e_1_2_1_19_1","doi-asserted-by":"publisher","DOI":"10.1038\/nm0996-1022"},{"key":"e_1_2_1_20_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1016113117851"},{"key":"e_1_2_1_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0142-9612(00)00196-4"},{"key":"e_1_2_1_22_1","doi-asserted-by":"publisher","DOI":"10.1002\/jbm.a.10165"},{"key":"e_1_2_1_23_1","unstructured":"Nektar Therapeutics.2003.PEG reagent catalog."},{"key":"e_1_2_1_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0169-409X(02)00022-4"},{"key":"e_1_2_1_25_1","volume-title":"The expanding role of mass spectrometry in biotechnology","author":"Siuzdak G","year":"2003"},{"key":"e_1_2_1_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0927-7757(99)00071-0"},{"key":"e_1_2_1_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-3659(01)00266-8"}],"container-title":["Biotechnology and Bioengineering"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fbit.21083","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/bit.21083","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T21:44:29Z","timestamp":1760046269000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/bit.21083"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2006,6,30]]},"references-count":26,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2006,12,20]]}},"alternative-id":["10.1002\/bit.21083"],"URL":"https:\/\/doi.org\/10.1002\/bit.21083","archive":["Portico"],"relation":{},"ISSN":["0006-3592","1097-0290"],"issn-type":[{"value":"0006-3592","type":"print"},{"value":"1097-0290","type":"electronic"}],"subject":[],"published":{"date-parts":[[2006,6,30]]}}}