This presentation accompanies the webinar which is available at: https://www1.gotomeeting.com/register/855617145 The AssayCheX process control panel is designed to provide assurance of the validity of each sample’s results for immunoassays performed using Luminex xMAP technology. The webinar covers the following topics: 1. AssayCheXTM Process Control Panel: an introduction 2. The product configuration 3. Functionality of each microsphere in process control panel 4. How to use AssayCheX along with current assays 5. Value provided by AssayCheX process control panel 6. Data analysis with MasterPlex® QT AssayCheX plug-in For more information, please visit: http://www.miraibio.com/reagents/assaychex-process-controls.html
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1. AssayCheX TMProcess Control Panel for Luminex/Bio-Plex
Immunoassays: Harnessing the true power ofxMAP technology
Presenters: Neeraja Venkateswaran, Ph.D. from Radix Biosolutions
Robert Lynde from MiraiBio Group of Hitachi Software
2. Webinar Recording
This presentation accompanies awebinar recordingwhich is
available at: https://www1.gotomeeting.com/register/855617145
3.
AssayCheX TMProcess Control Panel: an introduction
The product configuration
Functionality of each microsphere in process control panel
How to use AssayCheX along with current assays
Value provided by AssayCheX process control panel
Data analysis with MasterPlex QT AssayCheX plug-in
Overview 4. Introduction
AssayCheX Process Control Panelis for use with Luminex xMAP
based immunoassays.
Two main components of xMAP technology are:
Flow based analyzer
Fluorescent microspheres.
5. Sandwich Immunoassay
In Sandwich immunoassays the capture antibodies to analyte are
immobilized on microspheres and biotinylated detection antibody is
used in combination with streptavidin-phycoerythrin (SAPE) for
detecting the specific analyte.
6. 4-plex Process Control Panel
The AssayCheX process control panel is designed to provide
assurance of the validity of each samples results for immunoassays
performed using Luminex xMAP technology.
It is a 4 Plex panel
7. The Product Configuration
Size: 0.5 mL/vial
Each vial designed for for 5 plates
It is a 4 plex panel with regions 97, 98, 99, 100
Available for non magnetic assays
Research use only
Supplied in brown vial to minimize exposure to light
Shipped cold.
8. Functionality of Each Microsphere Bead Region Name
Functionality 97 Instrument Control Monitors the Luminex analyzer
reporterfluorescence measurement 98 Fluorescent Reporter Control
Monitors the addition of the fluorescent reporter
streptavidin-phycoerythrin (SA-PE) reagent to the well 99 Biotin
Detector Control Monitors the addition of the biotin-labeled
detector antibody reagent to the well 100 Negative or Non-specific
Control Monitors the non-specific binding (if any) due to the
sample matrix 9. Immunoassays With AssayCheX.. 10. How to use ?
Its as easy to use as.
Perform Assay Acquire Data Analyze Data 1 Add AssayCheX
Microspheres 2 Add Regions To Template 3 Export to MasterPlex 11.
How to use with your assay
Add the AssayCheX process control panel microspheres to your
assays 1X microsphere mixture immediately prior to adding the
microspheres to the wells. Add one L (1 L) of AssayCheX
microspheres to the 1X microsphere mixture for each well to be
tested.
Perform assay procedure as per the assay kit protocol.
For data acquisition, select the AssayCheX microsphere regions
(namely 97, 98, 99, 100) in addition to the kit-specific regions in
an xMAP data acquisition/analysis software package.
Mix microspheres well before use by vortexing or pipetting up
and down.
12. Analysis Algorithm
This 4-plex panel utilizes two independent acceptance-metrics
to ensure that each step of the immunoassay is performed correctly
and that the instrumentation is operating properly at the time of
data acquisition.
Threshold value: For catastrophic reagent or instrument
failure
Acceptance range: Range of high and low acceptance values to
accommodate normal assay variance
To confirm the validity of each sample result during data
analysis, plate threshold values and acceptance criteria ranges
must be determined.
13. Plate Failure Data 14. Individual Well Failure 15. Using
with current Assays.. No AssayCheX With AssayCheX 16. Average
Percentage Recovery Percentage recovery #of two controls for 11
different analytes # using average MFI values of triplicate samples
Anal01 Anal02 Anal03 Anal04 Anal05 Anal06 Anal07 Anal08 Anal09
Anal10 Anal11 No AssayCheX control 1 % Recovery 105.66 82.38 89.52
93.20 70.32 72.56 107.18 92.88 184.30 105.01 91.47 control 2 %
Recovery 77.87 79.00 77.06 98.37 65.04 181.94 85.19 83.00 93.46
77.31 81.17 With AssayCheX control 1 % Recovery 86.66 80.55 96.80
91.24 82.59 104.75 81.60 87.18 75.07 85.07 88.89 control 2 %
Recovery 105.49 104.34 109.30 105.39 83.20 93.01 103.88 102.75
101.52 101.24 105.02 17. Value provided.
Verify individual data points
Confirm assay performance
Troubleshoot with greater ease
Instantly flag potential failures
Reduce the need for replicates
http:// www.assaychex.com/results.php
18. Value provided.
The Three Rs of AssayCheX
Remove outliers.
Reduce retests.
Reduce replicates.
How?
19. Value provided.
By Using AssayCheX and MasterPlex:A powerful combination.
Define and check for thresholds for each AssayCheX microsphere
region in every well.
Flag failed wells for review.
Automatically mark failed wells as outliers and exclude them
from further calculations.
Display results in an easy-to-read view and data table.
