Developments, Use and Detection of Labeled Beads for Immunoassay Development:Anti-SARS-CoV-2 antibodies detection as an example
In this study, we proposed an anti-SARS-CoV-2 antibodies detection method based on a flow-based sandwich immunoassay and the use of magnetic beads. This presentation only focuses on developing and evaluating beads with the optimization of the fluorescence detection condition.
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[00:00:01.110]Hello. My name is Kyla, a graduate student
from the chemistry department today.
[00:00:06.120]I could talk about the development use
and detection of the label to bees for
[00:00:10.200]the immunoassay development.
[00:00:12.090]And [inaudible] antibody
detection is an example of this.
[00:00:16.640]Here is the overview of my
presentation, SARS cov two,
[00:00:20.630]she's known as so COVID-19
is an abbreviation of the
severe acute respiratory
[00:00:26.060]Coronavirus COVID-19 has a
high transmission lead and a
[00:00:31.190]long incubation period, allowing it to
spread quickly open without symptoms.
[00:00:36.020]These characteristics of the virus made
it hard to mitigate this pandemic will
[00:00:40.370]help this situation. There is a need
of the lipid accurate, affordable,
[00:00:44.960]and simple to use potable
taction device. In this study,
[00:00:49.130]we propose the portable detection
device based on the flow rates send each
[00:00:53.090]immunoassay and magnetic beads.
[00:00:55.880]But this presentation only focus on the
development and evaluation of the MES
[00:01:00.620]with the optimization of
the detection condition.
[00:01:03.620]There are some of the scheme
of the flow rates and HSE.
[00:01:07.940]These consists of the three steps first
combined sample with the capturing
[00:01:12.770]agent on the support for the
COVID-19 detections are scoped to
[00:01:17.570]spike a protein. As one has been
used as a capturing agent on support.
[00:01:23.270]Second at the secondary level,
the entire bodies. In our case,
[00:01:27.140]we use the sense dye for the labeling.
[00:01:29.600]Third Ditech labeled
antibiotics on the support.
[00:01:33.410]The goal of this study is to
develop you check the label,
[00:01:36.410]the 4k mobilize the magnetic bees for
the entire SARS cov empire bodies.
[00:01:40.940]Even when they say development,
[00:01:42.980]several factors need to be
concerned to achieve the goal.
[00:01:50.540]These factors preparation
piece responses of the label,
[00:01:54.390]the protein mobilize with, for
stability of the bees stability.
[00:01:58.190]The older bees immunoassay
application have been studied
[00:02:03.470]first target protein and mobilization
on the magnetic piece may need to
[00:02:08.150]immobilization method tested in here
if folksy and she put a base method
[00:02:13.230]as the graph describes a modified
Foxy method approved a higher protein
[00:02:17.930]contents after immobilization
different incubation time or so
[00:02:22.400]tested based on the
immobilized protein contents.
[00:02:25.820]The final coupling condition for the
protein will obliger and shim process was
[00:02:29.690]the modified the Foxy method with
the 48 hours incubation time.
[00:02:33.950]Next preparation of the label,
the protein for use of the label,
[00:02:37.490]the secondary interbodies
for the detection labeling.
[00:02:40.750]I have been chosen as we propose
the multianalyte detection,
[00:02:44.900]such as the different classes
of the COVID-19 anti-bodies IgG,
[00:02:49.580]IGA, IgM thighs, without interfering
each other need to be self-feed.
[00:02:54.680]Alexa dyes have been explicitly used
in the research area due to their,
[00:03:00.310]for different Alex [inaudible] had
chosen and use to label the secondary
[00:03:05.140]anti-bodies. And bovines from my Boomi,
[00:03:08.530]the amount of the immobilizer protein
on B's or dilatable the protein
[00:03:13.180]measured by using the BCSC
as the figure shown that a
[00:03:17.920]color change happens in
presence of the protein green to
[00:03:22.600]purple the amount of the protein
mobilize on the silica coordinator.
[00:03:26.200]Magnetic bees are the 16 to 20 milli, um,
[00:03:29.920]protein per gram of the bees
responses of the bees also tested
[00:03:35.110]eight in responses of the dial label.
[00:03:37.180]The protein mobilize RVs had been
obtained to correct the Cigna intensity of
[00:03:42.100]the bees with the different dyes,
Ty protein ratio and protein.
[00:03:46.120]The miles on the bees have been used to
normalize the signal intensity of the
[00:03:51.280]It was found that lately of similar
intensity decrease from the label dye with
[00:03:55.810]the roar excitation wavelengths to
higher excitation wavelengths next
[00:04:00.640]to presenter effect, Reggie
drew a sample in the mixture,
[00:04:04.150]several types of the buffer or the
actives had been evaluated to check the
[00:04:08.710]to remove the only acted a sample after
the mixing with the protein mobilize the
[00:04:12.490]bees 0.05% when 20 in pH PHP, 7.44,
[00:04:17.440]it proved that the lowest
rod value with the different
[00:04:22.300]label dates for stability of the bees,
[00:04:25.780]or is another important factor for the
validated bees based on the COVID-19 type
[00:04:31.750]they light red and blue light with
the use of the foil or evaluated
[00:04:37.150]the first figure obtained from the
use of the Le light without the foil.
[00:04:41.080]The second figure is for the use of the
blue light with the foil in the simple
[00:04:47.290]it is indicated that the use of the
foil helps with the dye stability.
[00:04:52.570]Let lie helps us AA of
the dye with the lower
[00:04:57.040]expectation we're labeling blue light.
[00:04:59.380]So with the labels at the
high excitation wavelengths,
[00:05:02.500]such as Alex's seven 15 here be
subtly in the portable itching
[00:05:07.390]of the bees have been
found at during this study,
[00:05:10.390]a greater decrease in signal
intensity of the fluorescence dye,
[00:05:14.140]following on increase the exposure
to light a decrease for assess
[00:05:18.970]intense steel corridor
in the use of the Fest,
[00:05:21.210]skim mold in the fluorescence detection.
[00:05:24.040]This is indicate that these
settling caused the decrease
of the signal intensity
[00:05:28.660]of the bees sample the stability Le label.
[00:05:32.210]The bees also tested the old and new
badges of the dilator with the protein
[00:05:36.580]mobilize. The bees represent a similar
responses. We got at least over the time.
[00:05:41.710]This represent that Cobell
conjugation between the vanity
[00:05:46.390]bees and the protein is stable or
where the time washing cycle and
[00:05:51.250]preachy f*g on the fluorescence intensity
over the bees were investigated the
[00:05:55.870]signal intensity of
bees over several cycle.
[00:05:59.210]The worst thing with a
pH 2.5, four or similar,
[00:06:04.400]these can be explained the possible
use of the bees in the multiple essay
[00:06:09.740]after testing or factors,
the prepare the label,
[00:06:12.570]the bees with the optimized conditions
such as a buffer or the tips were
[00:06:16.760]evaluated. Employer racing, the United
States, the left side of the graph,
[00:06:20.840]my presenter dilate with a protein
mobilize Amani bees can be suitable to
[00:06:25.550]serve as internal standard by giving
the a relatively consistent signal.
[00:06:29.810]The intensity's right side of the graph
shows the Montana light detection has
[00:06:34.640]seen Richard x-axis represented
the different samples and mix the,
[00:06:38.990]with the spec of the team mobilize
among the native bees and Alex a flow of
[00:06:42.600]four, four, eight, eight.
[00:06:43.670]They will the gold and
Ty human IgM served as a
[00:06:48.800]secondary entire bodies.
As it is shown here,
[00:06:52.460]the specificity responses between the
human IgM spiker protein sample with the
[00:06:56.780]secondary entire bodies obtain disprove
that the potential of the use of the
[00:07:02.230]the protein mobilize the magnetic bees
in murky anolyte detection for the entire
[00:07:06.950]COVID-19 anti-bodies here as a conclusion,
[00:07:11.180]target protein mobilized. Am I going to
be having prepared by a proxy method?
[00:07:15.890]They enable the protein mobilize.
[00:07:17.330]The magnetic bees have shown a linear
skin to intercity and a good stability.
[00:07:22.520]Diane labeled a protein mobilize among
ADP's that have ability to carry out
[00:07:27.110]multianalyte detection for the
anti SASCO anti-bodies detection.
[00:07:32.120]Thank you for all the support
from the Dr. Hage and then,
[00:07:36.410]and I tool, which is our collaborator
in this work. Thank you for listening.
[00:07:41.300]My presentation. I will have a
QA time on Thursday via zoom.
[00:07:44.990]I share the wrinkle in here. Thank you.
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