Biosensor for male infertility
A biosensor and identity technology, applied in the field of biosensors, can solve problems such as fertilization failure
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Embodiment 1
[0229] Example 1. Electrochemical biosensor for sperm binding using cyclic voltammetry
[0230] Fabrication of biosensors on screen-printed gold electrodes. Briefly, 10 µL of 2 mg / mL cysteamine hydrochloride was added to the gold electrode and dried at room temperature in the dark. The electrodes were washed with deionized water and then modified with 10 μL of citrate-capped gold nanoparticles. After drying and washing with deionized water, 10 μL of 50 mM nitrilotriacetic acid was added and incubated overnight at room temperature. It was subsequently blocked with 0.01 M phosphate buffer, pH 7.4, containing 1% bovine serum albumin. Add 10 µL of 10 mM nickel sulfate and incubate for 2 h at room temperature. After washing the electrode with 0.01 M phosphate buffer at pH 7.4, 0.5 μg of JUNO or ZP3 containing a polyhistidine tag was immobilized on the biosensor.
[0231] Semen samples were liquefied and diluted in semen preparation medium (#1070 / 1069, OrigioA / SDenmark, https:...
Embodiment 2
[0237] Example 2. Electrochemical biosensor for analysis of sperm binding using electrical impedance spectroscopy
[0238]Screen-printed gold electrodes were functionalized by 10 μl of 2 mg / ml cysteamine hydrochloride (CyHCl) and incubated for 2-4 h at room temperature in the dark. After washing the electrodes with deionized water, 5 μl of Asn-AuNPs were added onto the functionalized electrodes. The electrodes were incubated at room temperature for 5 h and then washed with deionized water. 5 μl of PB (10 mM, pH 7.4) containing 160 μg / ml JUNO and 160 μg / ml ZP3 in 5 μl was further added and incubated overnight at 4°C. Finally, the electrodes were blocked with 10 μl PBS containing 1% BSA for 5 h at 4°C.
[0239] Connect the modified electrode to the electrochemical workstation.
[0240] Add 5-50 µL of the pretreated semen sample to the working electrode, or dip the electrode into the sample vial. Sperm cells were allowed to bind to the modified electrodes for 1-10 minutes, ...
Embodiment 3
[0247] Example 3. Electrochemical biosensors for acrosome reaction
[0248] Screen-printed gold electrodes were functionalized by 10 μl of 2 mg / ml cysteamine hydrochloride (CyHCl) and incubated for 2-4 h at room temperature in the dark. After washing the electrodes with deionized water, 5 μl of Asn-AuNPs were added onto the functionalized electrodes. The electrodes were then incubated at room temperature for 5 h and then washed with deionized water. 5 μl of PB (10 mM, pH 7.4) containing 160 μg / ml JUNO and 160 μg / ml ZP3 in 5 μl was further added and incubated overnight at 4°C. Finally, the electrodes were blocked with 10 μl PBS containing 1% BSA for 5 h at 4°C.
[0249] Connect the modified electrode to the electrochemical workstation.
[0250] Add 5-50 µL of the pretreated semen sample to the working electrode, or dip the electrode into the sample vial. Sperm cells are bound to the modified electrodes while the binding is analyzed electrochemically by electrical impedanc...
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