Improving the reliability of human serum albumin-thiol group determination
Само за регистроване кориснике
2013
Аутори
Jovanović, Vesna B.Penezić-Romanjuk, Ana Z.
Pavićević, Ivan D.
Aćimović, Jelena M.
Mandić, Ljuba M.
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
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.
Кључне речи:
Albumin thiol group determination / Albumin isolation / Two-step precipitation method / Diabetes / Oxidative and carbonyl stressИзвор:
Analytical Biochemistry, 2013, 439, 1, 17-22Издавач:
- Academic Press Inc Elsevier Science, San Diego
Финансирање / пројекти:
- Алергени, антитела, ензими и мали физиолошки значајни молекули: дизајн, структура, функција и значај (RS-MESTD-Basic Research (BR or ON)-172049)
DOI: 10.1016/j.ab.2013.03.033
ISSN: 0003-2697
PubMed: 23583909
WoS: 000321029200006
Scopus: 2-s2.0-84877805283
Колекције
Институција/група
Hemijski fakultet / Faculty of ChemistryTY - JOUR AU - Jovanović, Vesna B. AU - Penezić-Romanjuk, Ana Z. AU - Pavićević, Ivan D. AU - Aćimović, Jelena M. AU - Mandić, Ljuba M. PY - 2013 UR - https://cherry.chem.bg.ac.rs/handle/123456789/1369 AB - 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. PB - Academic Press Inc Elsevier Science, San Diego T2 - Analytical Biochemistry T1 - Improving the reliability of human serum albumin-thiol group determination VL - 439 IS - 1 SP - 17 EP - 22 DO - 10.1016/j.ab.2013.03.033 ER -
@article{ author = "Jovanović, Vesna B. and Penezić-Romanjuk, Ana Z. and Pavićević, Ivan D. and Aćimović, Jelena M. and Mandić, Ljuba M.", year = "2013", 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.", publisher = "Academic Press Inc Elsevier Science, San Diego", journal = "Analytical Biochemistry", title = "Improving the reliability of human serum albumin-thiol group determination", volume = "439", number = "1", pages = "17-22", doi = "10.1016/j.ab.2013.03.033" }
Jovanović, V. B., Penezić-Romanjuk, A. Z., Pavićević, I. D., Aćimović, J. M.,& Mandić, L. M.. (2013). Improving the reliability of human serum albumin-thiol group determination. in Analytical Biochemistry Academic Press Inc Elsevier Science, San Diego., 439(1), 17-22. https://doi.org/10.1016/j.ab.2013.03.033
Jovanović VB, Penezić-Romanjuk AZ, Pavićević ID, Aćimović JM, Mandić LM. Improving the reliability of human serum albumin-thiol group determination. in Analytical Biochemistry. 2013;439(1):17-22. doi:10.1016/j.ab.2013.03.033 .
Jovanović, Vesna B., Penezić-Romanjuk, Ana Z., Pavićević, Ivan D., Aćimović, Jelena M., Mandić, Ljuba M., "Improving the reliability of human serum albumin-thiol group determination" in Analytical Biochemistry, 439, no. 1 (2013):17-22, https://doi.org/10.1016/j.ab.2013.03.033 . .