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Improving the reliability of human serum albumin-thiol group determination
dc.creator | Jovanović, Vesna B. | |
dc.creator | Penezić-Romanjuk, Ana Z. | |
dc.creator | Pavićević, Ivan D. | |
dc.creator | Aćimović, Jelena M. | |
dc.creator | Mandić, Ljuba M. | |
dc.date.accessioned | 2018-11-22T00:23:26Z | |
dc.date.available | 2018-11-22T00:23:26Z | |
dc.date.issued | 2013 | |
dc.identifier.issn | 0003-2697 | |
dc.identifier.uri | https://cherry.chem.bg.ac.rs/handle/123456789/1369 | |
dc.description.abstract | The thiol (Cys34) content of human serum albumin (HSA-SH) decreases during oxidative and carbonyl stress and, therefore, could represent a useful parameter in clinical practice. Nevertheless, the reliability of HSA-thiol determination with Ellman's method depends on the purity of isolated HSA. Determination of total serum thiols (mmol/L) and HSA-SH content (mmol -SH/mmol HSA) after HSA isolation from diabetic patient and control sera by a two-step precipitation with ammonium sulfate (AS), as well as HSA-SH contribution (%) to total serum thiols, was assessed. Purity and yield of isolated HSA were monitored spectrophotometrically and by native polyacrylamide gel electrophoresis. Precipitation of HSA from serum via a two-step method with AS produced HSA with 91.9 +/- 3.6% purity and 69.7 +/- 4.4% yield, allowing for precise (relative standard deviation of 3.2%) and reliable (comparing with total serum thiols) measurement of HSA-SH content with DTNB [5,5'-dithiobis-(2-nitrobenzoic acid)]. The content of the HSA-SH group in patients with type 2 diabetes was significantly (P lt 0.05) lower compared with that of the healthy cohort (0.483 +/- 0.067 vs. 0.561 +/- 0.054 mmol -SH/mmol HSA). Because the proposed method of HSA isolation is simple, time-efficient, and technically less demanding, and it also enables reliable determination of HSA-SH content, it is suitable for clinical practice. | en |
dc.publisher | Academic Press Inc Elsevier Science, San Diego | |
dc.relation | info:eu-repo/grantAgreement/MESTD/Basic Research (BR or ON)/172049/RS// | |
dc.rights | restrictedAccess | |
dc.source | Analytical Biochemistry | |
dc.subject | Albumin thiol group determination | en |
dc.subject | Albumin isolation | en |
dc.subject | Two-step precipitation method | en |
dc.subject | Diabetes | en |
dc.subject | Oxidative and carbonyl stress | en |
dc.title | Improving the reliability of human serum albumin-thiol group determination | en |
dc.type | article | |
dc.rights.license | ARR | |
dcterms.abstract | Павицевиц, Иван Д.; Пенезиц-Ромањук, Aна З.; Мандић, Љуба М.; Јовановић, Весна; Aћимовић, Јелена; | |
dc.citation.volume | 439 | |
dc.citation.issue | 1 | |
dc.citation.spage | 17 | |
dc.citation.epage | 22 | |
dc.identifier.wos | 000321029200006 | |
dc.identifier.doi | 10.1016/j.ab.2013.03.033 | |
dc.citation.other | 439(1): 17-22 | |
dc.citation.rank | M22 | |
dc.identifier.pmid | 23583909 | |
dc.type.version | publishedVersion | en |
dc.identifier.scopus | 2-s2.0-84877805283 |