دورية أكاديمية
أعرض في EDS

A high stable sample loading for analysis of adult alpha-thalassemia via the improved microarray isoelectric focusing of Hb species.

التفاصيل البيبلوغرافية
العنوان: A high stable sample loading for analysis of adult alpha-thalassemia via the improved microarray isoelectric focusing of Hb species.
المؤلفون: Sani A; School of Sensing Science and Engineering, School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, Shanghai 200240, China., Tian Y; School of Sensing Science and Engineering, School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, Shanghai 200240, China., Zha G; School of Sensing Science and Engineering, School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, Shanghai 200240, China., Xiao X; NHC Key Laboratory of Thalassemia Medicine, Key Laboratory of Thalassemia Medicine, Chinese Academy of Medical Sciences, Guangxi Key Laboratory of Thalassemia Research, Guangxi Medical University, Nanning 530021, PR China., Shah S; School of Sensing Science and Engineering, School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, Shanghai 200240, China., Idrees Khan M; School of Sensing Science and Engineering, School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, Shanghai 200240, China., Fan L; Student Innovation Center, Shanghai Jiao Tong University, Shanghai 200240, China., Zhang Q; School of Sensing Science and Engineering, School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, Shanghai 200240, China., Liu W; School of Sensing Science and Engineering, School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, Shanghai 200240, China., Wang Y; School of Physics and Astronomy, Shanghai Jiao Tong University, Shanghai 200240, China. Electronic address: wyx75@sjtu.edu.cn., Chen P; NHC Key Laboratory of Thalassemia Medicine, Key Laboratory of Thalassemia Medicine, Chinese Academy of Medical Sciences, Guangxi Key Laboratory of Thalassemia Research, Guangxi Medical University, Nanning 530021, PR China. Electronic address: cping62@hotmail.com., Cao C; School of Sensing Science and Engineering, School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, Shanghai 200240, China. Electronic address: cxcao@sjtu.edu.cn.
المصدر: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences [J Chromatogr B Analyt Technol Biomed Life Sci] 2024 Aug 15; Vol. 1244, pp. 124238. Date of Electronic Publication: 2024 Jul 14.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Elsevier Country of Publication: Netherlands NLM ID: 101139554 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1873-376X (Electronic) Linking ISSN: 15700232 NLM ISO Abbreviation: J Chromatogr B Analyt Technol Biomed Life Sci Subsets: MEDLINE
أسماء مطبوعة: Original Publication: Amsterdam ; New York : Elsevier, c2002-
مواضيع طبية MeSH: Isoelectric Focusing*/methods , alpha-Thalassemia*/blood, Humans ; Hemoglobins, Abnormal/analysis ; Hemoglobins, Abnormal/chemistry ; Adult ; Linear Models ; Reproducibility of Results ; Limit of Detection
مستخلص: The isoelectric focusing has realized various improvements, including the protocols and creation of mIEF (microcolumn isoelectric focusing) instruments with excellent sensitivity for screening of diabetes and beta thalassemia. However, the problem of manual sample loading and hydration for the mIEF limits the operational capacity for stably detecting and quantitating most abnormal hemoglobin (Hb). Herein, we provided a high stable sample loading protocol for analysis of alpha thalassemia and Hb variants. In contrast to the previous volume of 20 μl, a 100 µl blood sample solution in this protocol was optimized with mixture of 6.4-7.5 and 3-10 pH carrier ampholytes, pI markers and loaded for 30 mins IPG microcolumn hydration. The hydrated microcolumn was then automatically loaded onto the mIEF chip array to which CH 3 COOH and NH 4 OH act as anodic and cathodic solutions. Lastly, the IEF was run for 9 mins. Hb H, Barts, A 1c , F, A 2 and CS were simultaneously separated and focused with higher resolution and sensitivity in quantifying H and Barts as low as 0.6 and 0.5 % respectively. Accordingly, there was an enhanced stability and linearity with a rapid assay time of 45 secs per sample. Moreover, analysis showed a fitting linear relationship with conventional technology at R 2  = 0.9803 for H and R 2  = 0.9728 for Barts thereby indicating greater accuracy confirmed by the AUC. Hence, the developed protocol could simply be employed for high stable and throughput batch sample loading of hydration, and accurate separation and quantitation of Hb variants for alpha and beta thalassemia.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 Elsevier B.V. All rights reserved.)
فهرسة مساهمة: Keywords: Alpha thalassemia; Hemoglobin; Isoelectric focusing; Protocol; Quantitation
المشرفين على المادة: 0 (Hemoglobins, Abnormal)
تواريخ الأحداث: Date Created: 20240717 Date Completed: 20240808 Latest Revision: 20240808
رمز التحديث: 20240809
DOI: 10.1016/j.jchromb.2024.124238
PMID: 39018990
قاعدة البيانات: MEDLINE
الوصف
تدمد:1873-376X
DOI:10.1016/j.jchromb.2024.124238