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Systemic VEGF measurements are important in view of the most appropriate Anti-VEGF drug to be used for the treatment of acute retinopathy of prematurity (ROP). Cord blood samples from preterm (n\u2009=\u20096) infants, blood samples from preterm infants with treatment requiring ROP (n\u2009=\u200912), and blood samples from healthy adults (n\u2009=\u200910) were collected. Serum, citrate plasma, and serum from recalcified citrate blood were obtained. Levels of VEGF-A and platelet factor-4 (PF-4) were quantified by ELISA or AlphaLISA immunoassay. VEGF-A levels could be detected by both assays, with the AlphaLISA generating slightly lower levels in healthy adults, but not in cord blood of preterm infants. In plasma samples, VEGF levels ranged from non detectable to 181\u2009pg\/ml. PF-4 concentrations were between 0.16\u20133.88\u2009\u00b5g\/ml. Values of VEGF-A and PF-4 in serum and recalcified\u00a0serum were significantly higher compared to plasma through the release of these cytokines after platelet activation. In plasma samples of infants with ROP, VEGF-A could always be detected and its values ranged from 19.50 to 245.91\u2009pg\/ml and PF-4 concentrations were between 0.1 and 3.3\u2009\u00b5g\/ml. Using the AlphaLISA kit, we were able to detect VEGF in small sample volumes (5\u2009\u00b5l plasma or serum\/well) in premature infants with treatment requiring ROP and to monitor platelet activation by PF-4 detection. Minimal blood probe volumes reduce phlebotomy losses avoiding the risk of iatrogenic anemia, thus allowing close monitoring of the cytokine levels in these very fragile infants.<\/jats:p>","DOI":"10.1038\/s41598-019-43108-7","type":"journal-article","created":{"date-parts":[[2019,5,1]],"date-time":"2019-05-01T10:02:54Z","timestamp":1556704974000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Optimizing Measurement of Vascular Endothelial Growth Factor in Small Blood Samples of Premature Infants"],"prefix":"10.1038","volume":"9","author":[{"given":"Claudia C.","family":"Lopez Yomayuza","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Klaus T.","family":"Preissner","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Birgit","family":"Lorenz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Knut","family":"Stieger","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2019,5,1]]},"reference":[{"key":"43108_CR1","doi-asserted-by":"publisher","first-page":"67","DOI":"10.1016\/j.preteyeres.2015.06.002","volume":"49","author":"PA Campochiaro","year":"2015","unstructured":"Campochiaro, P. 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