Next Article in Journal
Trametes versicolor (Synn. Coriolus versicolor) Polysaccharides in Cancer Therapy: Targets and Efficacy
Previous Article in Journal
Brassicasterol with Dual Anti-Infective Properties against HSV-1 and Mycobacterium tuberculosis, and Cardiovascular Protective Effect: Nonclinical In Vitro and In Silico Assessments
Previous Article in Special Issue
Non-Invasive Delivery of Therapeutics into the Brain: The Potential of Aptamers for Targeted Delivery
Open AccessArticle

Evaluation of Aptamers as Affinity Reagents for an Enhancement of SRM-Based Detection of Low-Abundance Proteins in Blood Plasma

1
Institute of Biomedical Chemistry, Moscow 119121, Russia
2
Russian Scientific Center of Roentgenoradiology, Moscow 117485, Russia
*
Author to whom correspondence should be addressed.
Biomedicines 2020, 8(5), 133; https://doi.org/10.3390/biomedicines8050133
Received: 26 April 2020 / Revised: 18 May 2020 / Accepted: 22 May 2020 / Published: 24 May 2020
(This article belongs to the Special Issue Engineering Aptamers for Biomedical Applications II)
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. View Full-Text
Keywords: selected reaction monitoring; protein detection; blood plasma; SMAD4; aptamer-based affinity enrichment selected reaction monitoring; protein detection; blood plasma; SMAD4; aptamer-based affinity enrichment
Show Figures

Figure 1

MDPI and ACS Style

Radko, S.; Ptitsyn, K.; Novikova, S.; Kiseleva, Y.; Moysa, A.; Kurbatov, L.; Mannanova, M.; Zgoda, V.; Ponomarenko, E.; Lisitsa, A.; Archakov, A. Evaluation of Aptamers as Affinity Reagents for an Enhancement of SRM-Based Detection of Low-Abundance Proteins in Blood Plasma. Biomedicines 2020, 8, 133.

Show more citation formats Show less citations formats
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Search more from Scilit
 
Search
Back to TopTop