AssayCheX Process Control Panel for Luminex/Bio-Plex Immunoassays

AssayCheX TM Process Control Panel for Luminex/Bio-Plex Immunoassays: Harnessing the true power of xMAP technology Presenters: Neeraja Venkateswaran, Ph.D. from Radix Biosolutions Robert Lynde from MiraiBio Group of Hitachi Software

Webinar Recording
This presentation accompanies a webinar recording which is available at: https://www1.gotomeeting.com/register/855617145

AssayCheX TM Process 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

Introduction
AssayCheX™ Process Control Panel is for use with Luminex xMAP based immunoassays.
Two main components of xMAP technology are:
Flow based analyzer
Fluorescent microspheres.

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.

4-plex Process Control Panel
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.
It is a 4 Plex panel

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.

Functionality of Each Microsphere Bead Region Name Functionality 97 Instrument Control Monitors the Luminex analyzer reporter fluorescence 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

Immunoassays With AssayCheX..

How to use ?
It’s as easy to use as….
Perform Assay Acquire Data Analyze Data 1 Add AssayCheX Microspheres 2 Add Regions To Template 3 Export to MasterPlex

How to use with your assay
Add the AssayCheX process control panel microspheres to your assay’s 1X microsphere mixture immediately prior to adding the microspheres to the wells. Add one µL (1 µL) 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.

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.

Plate Failure Data

Individual Well Failure

Using with current Assays.. No AssayCheX With AssayCheX

Average Percentage Recovery Percentage recovery # of two controls for 11 different analytes # using average MFI values of triplicate samples Anal 01 Anal 02 Anal 03 Anal 04 Anal 05 Anal 06 Anal 07 Anal 08 Anal 09 Anal 10 Anal 11 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

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

Value provided….
The Three Rs of AssayCheX
Remove outliers.
Reduce retests.
Reduce replicates.
How?

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.
Automatically generate a final report of results.
http:// www.assaychex.com/value.php

Resources
assaychex.com
Detailed product information
miraibio.com
Online Store or sales@miraibio.com
AssayChex – 5 plates for $495 through the end of November
FREE Trial download of the MasterPlex software
Blog , Live chat, flash movies, tutorials, user community etc.
[email_address]
radixbiosolutions.com
Custom assay development
Custom production
Custom reagent production
Modified microspheres
[email_address]

AssayCheX Process Control Panel for Luminex/Bio-Plex Immunoassays

by MiraiBio Group of Hitachi Solutions America, Ltd. on Oct 30, 2009

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AssayCheX Process Control Panel for Luminex/Bio-Plex Immunoassays — Presentation Transcript