Fluorescence assay of non-transferrin-bound iron in thalassemic sera using bacterial siderophore.

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dc.contributor.author Sharma, Manisha en
dc.contributor.author Saxena, R en
dc.contributor.author Gohil, NK en
dc.coverage.spatial United States en
dc.date.accessioned 2012-02-23T01:18:24Z en
dc.date.issued 2009-11-15 en
dc.identifier.citation Analytical Biochemistry 394(2):186-191 15 Nov 2009 en
dc.identifier.uri http://hdl.handle.net/2292/11768 en
dc.description.abstract Transfusional iron overload associated with thalassemia leads to the appearance of non-transferrin-bound iron (NTBI) in blood that is toxic and causes morbidity and mortality via tissue damage. Hence, a highly sensitive and accurate assay of NTBI, with broad clinical application in both diagnosis and validation of treatment regimens for iron overload, is important. An assay based on iron chelation by a high-affinity siderophore, azotobactin, has been developed. The steps consist of blocking of native apotransferrin iron binding sites, mobilization of NTBI, ultrafiltration of all serum proteins, and finally the addition of the probe, which has a chromophore that fluoresces at 490 nm. Binding of Fe3+ to azotobactin quenches the fluorescence in a concentration-dependent manner. Measured NTBI levels in 63 sera ranged from 0.07 to 3.24 microM (0.375+/-0.028 microM [means+/-SEM]). It correlated well with serum iron and percentage transferrin saturation but not with serum ferritin. Pearson's correlation coefficients were found to be 0.6074 (P<0.0001) and 0.6102 (P<0.0001) for percentage transferrin saturation and total serum iron, respectively. The low values are due to the patients being under regular chelation therapy even prior to sampling, indicating that the method is sensitive to very low levels of NTBI, allowing a much lower detection limit than the available methods. en
dc.language eng en
dc.publisher Academic Press en
dc.relation.ispartofseries Analytical Biochemistry en
dc.rights Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. Details obtained from: http://www.sherpa.ac.uk/romeo/issn/0003-2697/ en
dc.rights.uri https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm en
dc.subject Adolescent en
dc.subject Adult en
dc.subject Apoproteins en
dc.subject Azotobacter vinelandii en
dc.subject Calibration en
dc.subject Child en
dc.subject Child, Preschool en
dc.subject Feasibility Studies en
dc.subject Female en
dc.subject Fluorescence en
dc.subject Humans en
dc.subject India en
dc.subject Iron en
dc.subject Male en
dc.subject Siderophores en
dc.subject Spectrometry, Fluorescence en
dc.subject Transferrin en
dc.subject Young Adult en
dc.subject beta-Thalassemia en
dc.title Fluorescence assay of non-transferrin-bound iron in thalassemic sera using bacterial siderophore. en
dc.type Journal Article en
dc.identifier.doi 10.1016/j.ab.2009.07.028 en
pubs.issue 2 en
pubs.begin-page 186 en
pubs.volume 394 en
dc.rights.holder Copyright: Academic Press en
dc.identifier.pmid 19632191 en
pubs.end-page 191 en
dc.rights.accessrights http://purl.org/eprint/accessRights/RestrictedAccess en
pubs.subtype Article en
pubs.elements-id 233827 en
pubs.org-id Medical and Health Sciences en
pubs.org-id Pharmacy en
dc.identifier.eissn 1096-0309 en
dc.identifier.pii S0003-2697(09)00514-4 en
pubs.record-created-at-source-date 2012-02-17 en
pubs.dimensions-id 19632191 en

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