Dose Response Curve

As described in our Bioassay Analysis with the Five Parameter Logistic (5-PL) Non-Linear Regression Curve-Fitting Model blog post, the C parameter in both the 4-PL and 5-PL models represent the concentration value for the inflection point of the curve. Depending on whether you have an inhibition or activation assay, the C parameter also represents the half maximal inhibitory concentration (IC50) or the half maximal effective concentration (EC50) respectively.

Anatomy of the 4 or 5 Parameter Logistic Model

MasterPlex ReaderFit offers full support for the 4 Parameter Logistic (4-PL) and 5 Parameter Logistic (5-PL) model equations making it an ideal tool for automated fitting of dose response curves and calculating the EC50 or IC50 values.

Here is a Quick Guide for this procedure:

1. To acquire MasterPlex ReaderFit, download and install the free 14-day trial.
2. If you have not yet imported your data, please refer to the community post on importing data into MasterPlex ReaderFit.
3. This is a sample dose response data set which contains 3 different sets of data from which we will generate 3 dose response curves (each outlined with a different color). Each curve set contains 8 data points with duplicates that are side-by-side.
   

   Sample Dose Response Data

4. Select 1 entire set of data points (including replicates) and click the blue Standard button. Please note that you can hold down the CTRL button and left click non-contiguous wells.
   

   Marking Data Sets

5. The Auto Fill dialog will pop up. This feature is only helpful if you have a serial dilution for your drug concentrations. If not, you will have to enter them in manually.
   

   Auto Fill Feature for Entering in Serial Dilution Concentrations

   In this example, Drug ABC was serially diluted from top (20,000 pg/mL) to bottom (0.26 pg/mL) with a dilution factor of 5.

6. Click Apply & View to preview the values in plate tab to confirm.
7. Repeat steps 2-4 for any other data sets you may have. In this example, there are 2 other data sets.
   For each cell, you will notice 2 numbers: the response value on the top and the drug dose on the bottom.
8. Click on the Standard Curve tab.
   
9. Make sure the Use Best Fit feature checkbox is unchecked and select the model equation (4-PL or 5-PL) you would like to use for the curve fit and then click Calculate.
   
   

   Model Equation Selection and Calculation

10. MasterPlex ReaderFit will then draw the dose response curve(s) according to the parameters you have chosen.
    
    

    Dose Response Curves based on the 4 Parameter Logistic

11. To view the EC50 or IC50 calculations, click on the individual curves (if you have more than one) and look for the C parameter listed above the dose response curve. In this case, this value represents the EC50 because this was an activation assay.
    
    

    EC50 value

For more information or to download the free 14-day trial, please visit the MasterPlex ReaderFit product page.

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As you will see, some of these are capable of outputting some pretty comprehensive statistics such as the lower limit of detection (llod) and conditional formatting where wells that exceed certain thresholds will be marked for you automatically. To view the sample screenshots of each report, click on the Details button.

Please refer to our community forum for detailed instructions on using the Export Manager.

If you would like to have a custom report template created for you, we can provide it as a service.  Please feel free to contact us at info@miraibio.com for details.

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The webinar covers the following topics:

1. AssayCheX^TM 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

Download the PowerPoint Presentation that accompanies this webinar.

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Learn how you can add a 4-plex Process control panel to any xMAP based immunoassay to evaluate every single well on your plate and save costs by reducing replicates and sample reruns. AssayChex gives you the power to:

• Verify individual data points
• Confirm assay performance
• Troubleshoot with greater ease
• Instantly flag potential failures
• Reduce the need for replicates

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For organizations working under the FDA 21 CFR Part 11 regulations, this plug-in offers:

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• MasterPlex ReaderFit 101
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• The Wait is Over: A New QC Tool for Luminex/Bio-Plex Analysis
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• AssayCheX Process Control Panel for Luminex/Bio-Plex Immunoassays
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• 10 Tips for MasterPlex QT v4.0 Power Users
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• Microplate Reader/Elisa Data Analysis made easy with MasterPlex ReaderFit
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• 10 Tips for MasterPlex QT v4.0 Power Users on August 20th at 11AM – 12PM PDT – Reserve your seat!
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If you cannot make the time for the MasterPlex ReaderFit webinar, you can view a previously recorded video of it and obtain the PowerPoint presentation slides.

A recording of the MasterPlex QT webinar will be available online after August 20th, 2009.

1. File Import Wizard
2. Automatic Detection of Outliers
3. Marking Wells and Grouping of Replicates
4. Entering Standard Information
5. Best Fit and Best Fit Parameters
6. Plotting Unknown Data Points on the Standard Curve
7. WYSIWYG Data Table
8. Chart Customization
9. Templates
10. Custom Reports
11. Virtual Plate

Download the PowerPoint Presentation if you would like to follow along with the recording.

Please click here to view the recorded MasterPlex ReaderFit webinar. Be sure to prepare some popcorn as the running time is about 40 minutes. Enjoy the show =)

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