{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,16]],"date-time":"2025-10-16T20:14:26Z","timestamp":1760645666399,"version":"build-2065373602"},"reference-count":48,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2009,4,16]],"date-time":"2009-04-16T00:00:00Z","timestamp":1239840000000},"content-version":"vor","delay-in-days":15,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Electrophoresis"],"published-print":{"date-parts":[[2009,4]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The vascular endothelial growth factor (VEGF) is involved in different sicknesses (cardiovascular diseases, cancer, and other). Out of the many components of the VEGF family, the A splice variant with 165 amino acids (VEGF<jats:sub>165<\/jats:sub>) is the main component. In spite of the potential as biomarker that this protein has, information about its physico\u2010chemical characteristics is scarce. In this study CIEF and MALDI\u2010TOF\u2010MS methods for intact recombinant human VEGF<jats:sub>165<\/jats:sub> are developed and applied to analyze this glycoprotein expressed in glycosylating (<jats:italic>Sf 21<\/jats:italic> insect cells) and non\u2010glycosylating (<jats:italic>Escherichia coli<\/jats:italic>) systems. Different parameters influencing the CIEF separation were studied. The developed CIEF method allowed for the separation of up to seven peaks in the VEGF<jats:sub>165<\/jats:sub> expressed in insect cells and up to three in VEGF<jats:sub>165<\/jats:sub> expressed in <jats:italic>E. coli<\/jats:italic>. The use of the presented method permits the estimation of the apparent p<jats:italic>I<\/jats:italic> of the different forms of VEGF<jats:sub>165<\/jats:sub> expressed in insect cells to be in a range of 6.8\u20138.2. The three peaks with intermediate p<jats:italic>I<\/jats:italic> values are observed in the protein expressed in both systems, insect cells and <jats:italic>E. coli<\/jats:italic>. The MALDI\u2010TOF\u2010MS method enabled to a rapid partial characterization of VEGF<jats:sub>165<\/jats:sub> based on its MS fingerprint. MALDI\u2010MS analysis of VEGF<jats:sub>165<\/jats:sub> expressed in insect cells shows the presence of, at least, four forms or groups of forms of VEGF<jats:sub>165<\/jats:sub> as a result of the different PTMs of the protein. According to the MALDI\u2010MS analysis, VEGF<jats:sub>165<\/jats:sub> expressed in <jats:italic>E. coli<\/jats:italic> was produced as a very homogeneous protein, although the results suggest the existence of some PTMs in the protein. The patterns of VEGF<jats:sub>165<\/jats:sub> of both origins obtained by CIEF and MALDI\u2010MS indicate the possibility of using these analytical methods to compare samples from people with different pathophysiological conditions. This work is thus a starting point to make possible the study of the role of the various forms of VEGF<jats:sub>165<\/jats:sub> as biomarkers. Finally, to the best of our knowledge, this is the first time that intact VEGF<jats:sub>165<\/jats:sub> has been analyzed by CIEF and MALDI\u2010TOF\u2010MS.<\/jats:p>","DOI":"10.1002\/elps.200800592","type":"journal-article","created":{"date-parts":[[2009,4,16]],"date-time":"2009-04-16T08:04:15Z","timestamp":1239869055000},"page":"1198-1205","source":"Crossref","is-referenced-by-count":7,"title":["CIEF and MALDI\u2010TOF\u2010MS methods for analyzing forms of the glycoprotein VEGF<sub>165<\/sub>"],"prefix":"10.1002","volume":"30","author":[{"given":"Sara","family":"Ongay","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Angel","family":"Puerta","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jose Carlos","family":"D\u00edez\u2010Masa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jonas","family":"Bergquist","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mercedes","family":"de Frutos","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2009,4,16]]},"reference":[{"key":"e_1_2_6_2_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.2479986"},{"key":"e_1_2_6_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/nm0603-669"},{"key":"e_1_2_6_4_2","doi-asserted-by":"publisher","DOI":"10.1210\/er.2003-0027"},{"key":"e_1_2_6_5_2","doi-asserted-by":"publisher","DOI":"10.1002\/jcb.240470305"},{"key":"e_1_2_6_6_2","doi-asserted-by":"publisher","DOI":"10.1080\/01674820601051034"},{"key":"e_1_2_6_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.pep.2005.09.023"},{"key":"e_1_2_6_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/0006-291X(91)91752-X"},{"key":"e_1_2_6_9_2","doi-asserted-by":"publisher","DOI":"10.1681\/ASN.2006010039"},{"key":"e_1_2_6_10_2","doi-asserted-by":"publisher","DOI":"10.1002\/ijc.22461"},{"key":"e_1_2_6_11_2","first-page":"2898","volume":"60","author":"Adams J.","year":"2000","journal-title":"Cancer Res."},{"key":"e_1_2_6_12_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0142(19960301)77:5<858::AID-CNCR8>3.0.CO;2-A"},{"key":"e_1_2_6_13_2","doi-asserted-by":"publisher","DOI":"10.1097\/00019501-200306000-00005"},{"key":"e_1_2_6_14_2","unstructured":"Elting J. J. Carney W. P. Hamer P. J. Patent WO200747955\u2010A2."},{"key":"e_1_2_6_15_2","first-page":"631","volume-title":"Capillary and Microchip Electrophoresis and Associated Microtechniques","author":"Girard M.","year":"2008"},{"key":"e_1_2_6_16_2","doi-asserted-by":"publisher","DOI":"10.1002\/elps.200600700"},{"key":"e_1_2_6_17_2","doi-asserted-by":"publisher","DOI":"10.1093\/ndt\/16.suppl_3.3"},{"key":"e_1_2_6_18_2","doi-asserted-by":"publisher","DOI":"10.1002\/elps.200800140"},{"key":"e_1_2_6_19_2","doi-asserted-by":"publisher","DOI":"10.1006\/abbi.1997.0145"},{"key":"e_1_2_6_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9673(98)00875-9"},{"key":"e_1_2_6_21_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1098-2787(1999)18:5<309::AID-MAS2>3.0.CO;2-L"},{"key":"e_1_2_6_22_2","doi-asserted-by":"publisher","DOI":"10.1002\/rcm.1239"},{"key":"e_1_2_6_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2004.04.021"},{"key":"e_1_2_6_24_2","doi-asserted-by":"publisher","DOI":"10.1002\/elps.200305831"},{"key":"e_1_2_6_25_2","doi-asserted-by":"publisher","DOI":"10.1002\/rcm.1290080215"},{"key":"e_1_2_6_26_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac960238z"},{"key":"e_1_2_6_27_2","doi-asserted-by":"crossref","first-page":"2288","DOI":"10.1093\/clinchem\/40.12.2288","volume":"40","author":"Hempe J. M.","year":"1994","journal-title":"Clin. Chem."},{"key":"e_1_2_6_28_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0003-2697(05)80014-4"},{"key":"e_1_2_6_29_2","doi-asserted-by":"publisher","DOI":"10.1002\/elps.1150110908"},{"key":"e_1_2_6_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/0021-9673(92)87041-6"},{"key":"e_1_2_6_31_2","doi-asserted-by":"publisher","DOI":"10.1002\/elps.200290005"},{"key":"e_1_2_6_32_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac961283"},{"key":"e_1_2_6_33_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9673(98)00326-4"},{"key":"e_1_2_6_34_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9673(02)00783-5"},{"key":"e_1_2_6_35_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0076-6879(96)70018-6"},{"key":"e_1_2_6_36_2","first-page":"453","volume":"830","author":"Rodr\u00edguez\u2010D\u00edaz R.","year":"1999","journal-title":"Electrophoresis"},{"key":"e_1_2_6_37_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0378-4347(96)00208-3"},{"key":"e_1_2_6_38_2","doi-asserted-by":"publisher","DOI":"10.1002\/elps.200600466"},{"key":"e_1_2_6_39_2","doi-asserted-by":"publisher","DOI":"10.1002\/elps.1150150105"},{"key":"e_1_2_6_40_2","first-page":"563","volume-title":"Capillary and Microchip Electrophoresis and Associated Microtechniques","author":"Wu J.","year":"2008"},{"key":"e_1_2_6_41_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chroma.2003.11.025"},{"key":"e_1_2_6_42_2","doi-asserted-by":"publisher","DOI":"10.1016\/0005-2795(71)90273-X"},{"key":"e_1_2_6_43_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.pep.2005.09.023"},{"key":"e_1_2_6_44_2","doi-asserted-by":"publisher","DOI":"10.1006\/prep.1995.1014"},{"key":"e_1_2_6_45_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-1176(97)00226-7"},{"key":"e_1_2_6_46_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0003-2670(00)83645-4"},{"key":"e_1_2_6_47_2","doi-asserted-by":"publisher","DOI":"10.1002\/rcm.1423"},{"key":"e_1_2_6_48_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac9604043"},{"key":"e_1_2_6_49_2","doi-asserted-by":"publisher","DOI":"10.1016\/0076-6879(91)98040-D"}],"container-title":["ELECTROPHORESIS"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Felps.200800592","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/elps.200800592","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T10:10:02Z","timestamp":1760523002000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/elps.200800592"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2009,4]]},"references-count":48,"journal-issue":{"issue":"7","published-print":{"date-parts":[[2009,4]]}},"alternative-id":["10.1002\/elps.200800592"],"URL":"https:\/\/doi.org\/10.1002\/elps.200800592","archive":["Portico"],"relation":{},"ISSN":["0173-0835","1522-2683"],"issn-type":[{"type":"print","value":"0173-0835"},{"type":"electronic","value":"1522-2683"}],"subject":[],"published":{"date-parts":[[2009,4]]}}}