Evaluation of Aptamers as Affinity Reagents for an Enhancement of SRM-Based Detection of Low-Abundance Proteins in Blood Plasma
| Autor: | K.G. Ptitsyn, Svetlana Novikova, Alexander Moysa, Elena A. Ponomarenko, Maria Mannanova, L. K. Kurbatov, Victor G. Zgoda, Y.Y. Kiseleva, Sergey P. Radko, Andrey Lisitsa, Alexander I. Archakov |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
medicine.drug_class Aptamer Medicine (miscellaneous) Monoclonal antibody 01 natural sciences SMAD4 General Biochemistry Genetics and Molecular Biology Article law.invention 03 medical and health sciences law Blood plasma aptamer-based affinity enrichment medicine lcsh:QH301-705.5 Chromatography biology Chemistry 010401 analytical chemistry Selected reaction monitoring 0104 chemical sciences 030104 developmental biology lcsh:Biology (General) Reagent selected reaction monitoring biology.protein Recombinant DNA Target protein Antibody human activities protein detection blood plasma |
| Zdroj: | Biomedicines Volume 8 Issue 5 Biomedicines, Vol 8, Iss 133, p 133 (2020) |
| ISSN: | 2227-9059 |
| Popis: | Selected reaction monitoring (SRM) is a mass spectrometric technique characterized by the exceptionally high selectivity and sensitivity of protein detection. However, even with this technique, the quantitative detection of low- and ultralow-abundance proteins in blood plasma, which is of great importance for the search and verification of novel protein disease markers, is a challenging task due to the immense dynamic range of protein abundance levels. One approach used to overcome this problem is the immunoaffinity enrichment of target proteins for SRM analysis, employing monoclonal antibodies. Aptamers appear as a promising alternative to antibodies for affinity enrichment. Here, using recombinant protein SMAD4 as a model target added at known concentrations to human blood plasma and SRM as a detection method, we investigated a relationship between the initial amount of the target protein and its amount in the fraction enriched with SMAD4 by an anti-SMAD4 DNA-aptamer immobilized on magnetic beads. It was found that the aptamer-based enrichment provided a 30-fold increase in the sensitivity of SRM detection of SMAD4. These results indicate that the aptamer-based affinity enrichment of target proteins can be successfully employed to improve quantitative detection of low-abundance proteins by SRM in undepleted human blood plasma. |
| Databáze: | OpenAIRE |
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