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. 2024 Jan 22;29(2):535.
doi: 10.3390/molecules29020535.

Design and Validation of a Short Novel Estradiol Aptamer and Exploration of Its Application in Sensor Technology

Hongyan Jin  1 , Yan Cheng  1 , Fanli Kong  2   3 , He Huang  1 , Zhenjun Yang  4 , Xinyi Wang  2   3 , Xinxia Cai  2   3 , Jinping Luo  2   3 , Tao Ming  5
Affiliations

Design and Validation of a Short Novel Estradiol Aptamer and Exploration of Its Application in Sensor Technology

Hongyan Jin et al. Molecules. .

Abstract

The specific and sensitive detection of 17β-estradiol (E2) is critical for diagnosing and treating numerous diseases, and aptamers have emerged as promising recognition probes for developing detection platforms. However, traditional long-sequence E2 aptamers have demonstrated limited clinical performance due to redundant structures that can affect their stability and recognition ability. There is thus an urgent need to further optimize the structure of the aptamer to build an effective detection platform for E2. In this work, we have designed a novel short aptamer that retains the key binding structure of traditional aptamers to E2 while eliminating the redundant structures. The proposed aptamer was evaluated for its binding properties using microscale thermophoresis, a gold nanoparticle-based colorimetric method, and electrochemical assays. Our results demonstrate that the proposed aptamer has excellent specific recognition ability for E2 and a high affinity with a dissociation constant of 92 nM. Moreover, the aptamer shows great potential as a recognition probe for constructing a highly specific and sensitive clinical estradiol detection platform. The aptamer-based electrochemical sensor enabled the detection of E2 with a linear range between 5 pg mL-1 and 10 ng mL-1 (R2 = 0.973), and the detection capability of a definite low concentration level was 5 pg mL-1 (S/N = 3). Overall, this novel aptamer holds great promise as a valuable tool for future studies on the role of E2 in various physiological and pathological processes and for developing sensitive and specific diagnostic assays for E2 detection in clinical applications.

Keywords: 17β-estradiol; aptamer; colorimetric; electrochemical; microscale thermophoresis.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
MST results of HEV1 binding with E2. (A) MST detection result of HEV1; (B) MST binding curve of HEV1.
Figure 2
Figure 2
(A) Visual observation results of different CNaCl; (B) absorption spectra of different CNaCl. (C) color development and detection spectra after adding different concentrations of M70; (D) color development and detection spectra after adding different concentrations of HEV1.
Figure 3
Figure 3
(A) Detecting E2 with M70. (A1) The actual detection diagram, (A2) the relationship diagram between the absorbance ratio A660/A520 of the system and CE2 and (A3) the linear regression curve. (B) Detecting E2 with HEV1. (B1) The actual detection diagram, (B2) the relationship diagram between the absorbance ratio A660/A520 of the system and CE2 and (B3) the linear regression curve.
Figure 4
Figure 4
(A) The SEM result of the modified electrode surface. (B) The TEM result of the synthesized nanocomposites.
Figure 5
Figure 5
(A) CV response of working electrodes subjected to different treatments. (B) DPV response of working electrodes subjected to different treatments.
Figure 6
Figure 6
(A) The DPV response of the M70 sensor when detecting different CE2; (B) M70 sensor detection result: calibration curve between DPV response and logarithmic CE2; (C) DPV response when HEV1 sensor detects different CE2; (D) HEV1 sensor detection result: calibration curve between DPV response and logarithmic CE2.
Figure 7
Figure 7
The schematic of short-aptamer modification of E2. (A) Scheme for Cutting M70 Adapters to Obtain HEV2; (B) Scheme for Cutting M70 Adapters to Obtain HEV1.
See this image and copyright information in PMC

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