Devan, Subramaniam-Betty Sheila and Ramli, Rosli and Alshehade, Salah Abdalrazak and Lim, Sharoen Yu Ming and Mamat, Noorhidayah (2025) Comparison of the techniques for isolating immunoassay-suitable proteins from heterogeneous fecal samples. Analytical Biochemistry, 698. p. 115748. ISSN 0003-2697, DOI https://doi.org/10.1016/j.ab.2024.115748.
Full text not available from this repository.Abstract
Immunoassays could provide valuable insights into disease biomarkers and gut health by measuring fecal proteins. However, reliably isolating intact proteins from feces is challenging due to its heterogeneous and variable composition. This paper aims to review and compare different methods for extracting proteins from fecal samples to make them suitable for immunoassay analysis. Mechanical homogenization helps release proteins by disrupting solids, while protease inhibitors preserve protein integrity. Detergents like SDS solubilize proteins by disrupting hydrophobic interactions. Organic solvents such as acetone precipitate proteins and remove contaminants. Thermal treatment denatures proteases. Immunocapture uses antibodies to purify target proteins away from interference selectively. Commercial kits contain optimized buffers but may be cost-prohibitive. Combining mechanical, chemical, and immunological techniques synergistically integrates their advantages, improving the recovery of native proteins with reduced matrix effects. While all methods have merits, tailored protocols integrating multiple mechanisms appear most promising for extracting immunoassay-suitable fecal proteins. Further optimization and standardization of such combination approaches matched to proteins and assays of interest helps expand noninvasive fecal proteome analysis.
| Item Type: | Article |
|---|---|
| Funders: | UNSPECIFIED |
| Uncontrolled Keywords: | Fecal protein extraction; Sample preparation; Immunoassay; Proteomics |
| Subjects: | Q Science > QH Natural history R Medicine > R Medicine (General) |
| Divisions: | Faculty of Medicine Faculty of Science > Institute of Biological Sciences |
| Depositing User: | Ms. Juhaida Abd Rahim |
| Date Deposited: | 04 Mar 2025 03:32 |
| Last Modified: | 04 Mar 2025 03:32 |
| URI: | http://eprints.um.edu.my/id/eprint/47794 |
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