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Isoelectric focusing array with immobilized pH gradient and dynamic scanning imaging for diabetes diagnosis. | LitMetric

Isoelectric focusing array with immobilized pH gradient and dynamic scanning imaging for diabetes diagnosis.

Anal Chim Acta

Department of Instrument Science and Engineering, Shanghai Engineering Research Center for Intelligent Diagnosis and Treatment Instrument, School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China; School of Life Sciences and Biotechnology, State Key Laboratory of Microbial Metabolism, Shanghai Jiao Tong University, Shanghai, 200240, China. Electronic address:

Published: July 2019

AI Article Synopsis

  • A novel isoelectric focusing (IEF) system was created using 24 immobilized pH gradient (IPG) columns to overcome the lengthy processes associated with traditional IEF, such as rehydration and staining.
  • The new system significantly reduced the time required for rehydration to just 25 minutes and IEF to 4 minutes, allowing for dynamic scanning of 24 columns in just 2 minutes.
  • The IEF array demonstrated good sensitivity and recovery rates, proving effective for separating and analyzing HbA, an important biomarker for diabetes diagnosis in blood samples.

Article Abstract

A traditional immobilized pH gradient (IPG) has a high stability for isoelectric focusing (IEF) but suffers from time-consuming rehydration, focusing and staining-imaging as well as complex performance. To address these issues, an IEF system with an array of 24 IPG columns (10 mm × 600 μm × 50 μm) and dynamic scanning imaging (DSI) was firstly designed for protein focusing. Moreover, two IPG columns (pH 4-9 and pH 6.7-7.7 of 10 mm in length) were firstly synthesized for IEF. A series of experiments were carried out based on the IEF array. In contrast to a traditional IPG IEF with more than 10 h rehydration, 5-14 h IEF and ca 10 h stain-imaging, the IEF array had the following merits: 25 min rehydration for sample loading, 4 min IEF, and 2 min dynamics scanning of 24 columns, well addressing the issues of traditional IEF. Furthermore, the IEF array had fair sensitivity (LOD of 60 ng), good recovery (95%), and high stability (1.02% RSD for intra-day and 2.16% for inter-day). Finally, the developed array was successfully used for separation and determination of HbA (a key biomarker for diabetes diagnosis) in blood samples. All these results indicated the applicability of the developed IEF array to diabetes diagnosis.

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Source
http://dx.doi.org/10.1016/j.aca.2019.03.014DOI Listing

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