11 Hormone Aptamers in Endocrine-Related Diseases
327
11.2.2 Progesterone-Related Aptamers
Progesterone (P4) is a 21-carbon steroid hormone secreted mainly by the corpus
luteum, and it is necessary for breast tissue development and pregnancy maintenance. Progesterone can regulate the menstrual cycle in women and plays a vital role
in menopausal hormone therapy and oral contraceptives [27]. The serum progesterone concentration of adult women is generally within the range of 0.48 –9.5 nM,
while the serum progesterone concentration of pregnant women can rise to 731 nM
[28]. The level of progesterone in the human body is extremely low, so the detection of progesterone has always been a problem. Various methods have been developed to detect progesterone such as immunoassays and chromatographic techniques.
The immunoassay methods include radioimmunoassay, ELISA, electrochemical
immunoassay. The chromatographic methods include high-performance liquid chromatography, liquid chromatography-mass spectrometry, gas chromatography-mass
spectrometry [29]. These methods necessitate more expensive equipment or tedious
complex processes due to the cross-reaction of antibodies and their batch-to-batch
variations [30].To overcome these problems, researchers have successfully generated
progesterone aptamer and developed several electrochemical aptasensors based on
different strategies.
In 2015, Contreras-Jimenez et al. [20] firstly obtained a P4 aptamer from a diverse
random library of ∼10
15 ss DNA sequences through in vitro selection procedures.
The best aptamer with an excellent KD of 17 nM was named P4G13, which shows
no cross-reactivity to analogs like E2 and Norethisterone (NET). P4G13 exhibited
a satisfying specific binding to P4 when the aptamer was hybridized with a 10mer short complementary sequence. Contreras-Jimenez also constructed a label-free
aptasensor to achieved a linear dynamic range from 10 to 60 ng/mL with a limit of
detection of 0.90 ng/mL of P4 in tap water samples. Skouridou et al. [31] further
explored the characteristic of P4G13. They found that the P4G13 aptamer also binds
with high affinity to 17α-hydroxyprogesterone, testosterone, and androstenedione
with KD of 3.5 nM, 31 nM, and 33 nM, respectively.
Alhadrami et al. [21] further truncated P4G13 into two different domains
(P4G13T1 and P4G13T2) based on the secondary structure of full-length aptamer
selected in their previous work [20]. They developed a fluorescence-based aptasensor
via competitive displacement assay for the detection of progesterone with high
affinity and specificity. The dissociation constant of the truncated aptamer-P4 was
enhanced by 16-fold compared to that of P4G13. The aptasensor based on the
truncated sequence P4G13T2 has shown high sensitivity with a detection limit of
110 pg/ml without significant cross-reactivity with similar target congeners, e.g., E2,
bisphenol A (BPA) and Vitamin D, the aptasensor also shown high recovery rates of
progesterone from spiked tap water and urine.
Lumei Wang group [32] further develop a colorimetric aptasensor based on fulllength P4G13 to detect P4 in human serum and urine (Fig. 11.3). This aptasensor
with high selectivity is designed based on the aggregation of gold nanoparticles
(AuNPs) controlled by the interactions among P4, P4 aptamer, and cationic surfactant
327
11.2.2 Progesterone-Related Aptamers
Progesterone (P4) is a 21-carbon steroid hormone secreted mainly by the corpus
luteum, and it is necessary for breast tissue development and pregnancy maintenance. Progesterone can regulate the menstrual cycle in women and plays a vital role
in menopausal hormone therapy and oral contraceptives [27]. The serum progesterone concentration of adult women is generally within the range of 0.48 –9.5 nM,
while the serum progesterone concentration of pregnant women can rise to 731 nM
[28]. The level of progesterone in the human body is extremely low, so the detection of progesterone has always been a problem. Various methods have been developed to detect progesterone such as immunoassays and chromatographic techniques.
The immunoassay methods include radioimmunoassay, ELISA, electrochemical
immunoassay. The chromatographic methods include high-performance liquid chromatography, liquid chromatography-mass spectrometry, gas chromatography-mass
spectrometry [29]. These methods necessitate more expensive equipment or tedious
complex processes due to the cross-reaction of antibodies and their batch-to-batch
variations [30].To overcome these problems, researchers have successfully generated
progesterone aptamer and developed several electrochemical aptasensors based on
different strategies.
In 2015, Contreras-Jimenez et al. [20] firstly obtained a P4 aptamer from a diverse
random library of ∼10
15 ss DNA sequences through in vitro selection procedures.
The best aptamer with an excellent KD of 17 nM was named P4G13, which shows
no cross-reactivity to analogs like E2 and Norethisterone (NET). P4G13 exhibited
a satisfying specific binding to P4 when the aptamer was hybridized with a 10mer short complementary sequence. Contreras-Jimenez also constructed a label-free
aptasensor to achieved a linear dynamic range from 10 to 60 ng/mL with a limit of
detection of 0.90 ng/mL of P4 in tap water samples. Skouridou et al. [31] further
explored the characteristic of P4G13. They found that the P4G13 aptamer also binds
with high affinity to 17α-hydroxyprogesterone, testosterone, and androstenedione
with KD of 3.5 nM, 31 nM, and 33 nM, respectively.
Alhadrami et al. [21] further truncated P4G13 into two different domains
(P4G13T1 and P4G13T2) based on the secondary structure of full-length aptamer
selected in their previous work [20]. They developed a fluorescence-based aptasensor
via competitive displacement assay for the detection of progesterone with high
affinity and specificity. The dissociation constant of the truncated aptamer-P4 was
enhanced by 16-fold compared to that of P4G13. The aptasensor based on the
truncated sequence P4G13T2 has shown high sensitivity with a detection limit of
110 pg/ml without significant cross-reactivity with similar target congeners, e.g., E2,
bisphenol A (BPA) and Vitamin D, the aptasensor also shown high recovery rates of
progesterone from spiked tap water and urine.
Lumei Wang group [32] further develop a colorimetric aptasensor based on fulllength P4G13 to detect P4 in human serum and urine (Fig. 11.3). This aptasensor
with high selectivity is designed based on the aggregation of gold nanoparticles
(AuNPs) controlled by the interactions among P4, P4 aptamer, and cationic surfactant