Mat Rasid, Siti Norain and Abd. Rahim, Nour El Huda and Mohamed Bakrim, Norbaiyah and A.Talib, Norlelawati and Abdull Jalil, Mohd Asyraf and Idorus, Mohd Yusri and Md Rosli, Ahmad Nabil (2023) Optimization of a two-dimensional electrophoresis protocol for plasma proteomic profiling of obese schizophrenia patients. In: 36th Malaysian Society of Pharmacology and Physiology Annual Scientific Meeting 2023, https://www.36thmspp2023.com/, Bangi Resort Hotel, Selangor.
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Abstract
The proteomic approach is particularly effective for studying the association between obesity and schizophrenia. It allows for a comprehensive analysis of the complete proteome, leading to substantial breakthroughs in biomarker discovery and drug development. Isoelectric focusing (IEF) and SDS-PAGE procedures are combined in the proteomic approach known as twodimensional electrophoresis (2-DE), which separates proteins according to their isoelectric point and mass. This study aimed to investigate optimized conditions for the 2-DE technique by focusing on the selection of an immobilized pH gradient (IPG) strip. Protein extraction was performed on pooled plasma samples from 10 obese schizophrenia patients. The extracted protein samples were loaded onto two different pH (7 cm) IPG strips. The pH ranges between (I) 3 – 10 and (ii) 4 – 7. IEF was conducted following the PROTEAN IEF Cell System protocol, followed by SDS-PAGE. The resulting gels were stained with BioSafe Coomassie stain and washed with milliQ water. The stained gels were scanned, and the images were analyzed using PD Quest software. High-abundance proteins with a molecular weight range of 60 – 80 kDa were detected on both IPG strips. The results showed that using a pH 3 – 10 IPG strip, 245 protein spots were detected and distributed throughout the gel, with a notable concentration in the middle. Whereas using a pH 4 – 7 IPG strip resulted in the detection of 321 protein spots, indicating a higher quantity of protein spots with increased intensity. This is attributed to the improved fractionation of proteins resulting from the narrower and more focused pH range. Thus, it can be inferred that utilizing this pH range will yield optimal outcomes in protein separation and analysis. This study suggests selecting a pH 4 – 7 IPG strip is the recommended choice to achieve enhanced resolution and precise detection of protein spots in plasma samples from obese schizophrenia patients when employing the 2-DE method.
| Item Type: | Conference or Workshop Item (Poster) |
|---|---|
| Uncontrolled Keywords: | Proteomics; 2-Dimensional Electrophoresis; IPG strip; Protein separation |
| Subjects: | Q Science > QP Physiology R Medicine > R Medicine (General) R Medicine > RC Internal medicine > RC321 Neuroscience. Biological psychiatry. Neuropsychiatry |
| Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button): | Kulliyyah of Medicine > Department of Basic Medical |
| Depositing User: | Dr Nour El Huda Abd Rahim |
| Date Deposited: | 25 Aug 2023 11:59 |
| Last Modified: | 25 Aug 2023 11:59 |
| URI: | http://irep.iium.edu.my/id/eprint/106200 |
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