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The Detection of Fertility Hormones via Antibody Immobilization and Electrochemical Impedance Spectroscopy. Student: Stephanie Maxwell Mentor: Dr. Jeffrey La Belle. The Objective:. WHY?. More than 500,000 pregnancies each year end in miscarriage (occurring during the first 20 weeks). - PowerPoint PPT Presentation
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1The Detection of Fertility Hormones via Antibody
Immobilization and Electrochemical Impedance
Spectroscopy
Student: Stephanie Maxwell Mentor: Dr. Jeffrey La Belle
2
The Objective:
3WHY?• More than 500,000 pregnancies each year end in
miscarriage (occurring during the first 20 weeks).• Over six million women in the United States struggle with
becoming or staying pregnant. • Hormone levels are constantly changing, and at times
drastically. • Pregnancies among women aged >40 years are associated
with more non-severe complications, more premature births, more congenital malformations and more interventions at birth.
4
Estradiol Molecule: Estradiol is a hormone associated with triggering responses that initiate ovulation
Progesterone Molecule: Progesterone is a steroid hormone, and it plays an important role in the preparation for and maintenance of pregnancies
Follicle Stimulating Molecule: The amount of FSH varies throughout a woman's menstrual cycle and is highest just before she releases an egg
Target Hormones
5
Methods: Immobilization
6Methods: Cyclic Voltammetry
0 2 4 6 8
-2
-1
0
1
2
Time (units)
Volta
ge (V
)
INPUT OUTPUT
7Methods: Electrochemical Impedance Spectroscopy
1 10 100 1000 10000 1000000
100
200
300
400
500
Frequency
Slo
pe (l
og)
1 10 100 1000 10000 1000000.0
0.2
0.4
0.6
0.8
Frequency
R-s
q (lo
g)
8Calibration Curve
-3 -2 -1 0 1 2 31060
1080
1100
1120
1140
1160
1180
1200
1220
1240
1260
f(x) = 20.9690174503249 x + 1194.13176366523R² = 0.996093830867349
Log[], log(ng/mL)
Impe
danc
e (o
hm)
Calibration curve created from the average of 7 different concentration gradients performed through EIS. Impedances were taken at the specific binding frequency (371Hz).
9Interferent Testing
-3 -2 -1 0 1 2 3 4 52000
2100
2200
2300
2400
2500
2600
P+ELinear (P+E)Linear (P+E)P+ALinear (P+A)PLinear (P)
Log [] pg/mL
Impe
denc
e (o
hm)
Calibration curves created after adding a concentration gradient of interferents to a concentration gradient of progesterone.
10Calibration in blood
0 10 20 30 40 50 60 70 80 90 1000
5
10
15
20
25
0
0.2
0.4
0.6
0.8
1
1.2slope
R2
% blood
slope R2
The effect of blood percentage on the slopes and R squared values of calibration curves of progesterone at 371Hz
11Conclusions • The optimal binding frequency of progesterone was found
(371Hz) and a linear relationship between impedance and log concentration was determined
• This relation is reproducible (R^2=0.99, n=7) • The response decreases with an increase in blood
percentage• The relation holds true with Estradiol acting as an
interferent; however a lot of noise is produced with the addition of Albumin
12Next steps• Non-target testing will also be completed to further prove specificity of the antibody
• Selectivity
• Sensitivity
• Combining and analyzing data from all targets
13
http://www.progesteronetherapy.com/image-files/graph.jpg
Electrochemical Impedance Spectroscopy
ν1ν2
ν3
Target Concentration
Impe
danc
e (Z
)
Simultaneous detection of
multiple hormones
associated with pregnancy
and/or fertility
ProgesteroneFSH
Estradiol
Final Goal
14
Thank you!