PQ Plate IRIS

PQ Plate IRIS

Introducing Mabtech's Performance Qualification (PQ) plate a fast, robust, and objective test to ensure that your IRIS or IRIS 2 performs according to its specifications. Our PQ plates offer a unique solution for verifying that the reader fulfills quality specifications and to ensure reproducibility, providing peace of mind knowing that experiments are accurate and reliable.

With the PQ plate, the Apex software conducts a series of quantitative and qualitative tests making it easy to identify any incorrect or inconsistent operation of the reader. A report is generated and the data is automatically saved and can be exported as a PDF for your records.

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Product specifications

Intended use

Serum/Plasma samples

Recommendation

Parameters tested: 

Light intensityVerification of stable light path from LED source to camera by collection of intensity data.
Focus

Determination of focus level to ensure consistent camera positioning and fixed focus.

RAWspot algorithmRAWspot algorithm assessed by collecting Spot Forming Units (SFU) and Relative Spot Volume (RSV) from PQ plate spots.

Shipping and Storage

Features

The PQ plate is analyzed with the PQ protocol in the Mabtech Apex software in a straightforward manner. Simply insert the PQ plate, select the PQ protocol, and the instrument will run the test and analyze the results. The data obtained is compared to baseline levels set for the PQ plate in the instrument. Each plate comes with a unique serial number, which is registered in Mabtech Apex, ensuring correct comparison. A report is automatically generated in non-modifiable pdf format, and the results are saved on the computer.


Following the test, a symbol indicating if the instrument passed is displayed in Mabtech Apex. Whenever logging into the instrument, the PQ symbol is clearly visible. If the test criteria were not fulfilled, you will receive a notification to contact us at Mabtech for support. 


We recommend that the qualification test is performed at regular intervals, and the software will let you know when it’s time to do so. When not in use, the PQ plate should be stored in its protective case in the dark, protected from dust, at room temperature.
 

Different sections of the PQ-plate:
1.    Tests focus and RAWspot algorithm in FluoroSpot
2.    Tests LED light intensity in FluoroSpot
3.    ELISpot test

Find out which analyte combinations we have evaluated in T cell FluoroSpot and which combinations are affected by capture effects or not. 

What is a capture effect?

When capture antibodies with different specificities are coated together, the capture of one cytokine may affect the secretion of other cytokines. This is usually more pronounced when studying T cell responses with polyclonal stimuli compared to antigen-specific responses. Capture effects are seldom a problem and can often be counteracted by the addition of an anti-CD28 antibody.

Depiction of capture effects observed in FluoroSpot

(1) IL-2 secreted by the activated T cell is captured by coated anti-IL-2 capture antibodies. (2) As a result, IL-2-stimulation of the T cell itself (autocrine stimulation) as well as nearby T cells (paracrine stimulation) is impaired, ultimately leading to (3) decreased IFN-γ secretion.

Co-stimulation with anti-CD28

Anti-CD28 mAb provides a co-stimulatory signal to antigen-specific responses by binding to CD28 on T cells. The addition of an anti-CD28 mAb to the cell culture enhances antigen-specific responses and can counteract capture effects. For example, the presence of IL-2 capture antibodies may result in reduced activation of T cells, as capturing of IL-2 decreases the amount of available IL-2 and thereby dampens the IL-2-mediated stimulation of T cells. The addition of anti-CD28 mAb restores IFN-γ responses (depicted in the below images). Further optimization may be necessary, depending on the cells and stimuli used. Too high concentrations of the anti-CD28 mAb may result in an elevation of non-specific cytokine secretion.

Overcoming capture effects using anti-CD28 anitbodies in FluoroSpot

(1) An anti-CD28 antibody can be added to provide a co-stimulatory signal that can restore (2) e.g. IFN-γ responses.

How to investigate capture effects

Our FluoroSpot Plus kits are evaluated for capture effects, and in studies of T cell responses we recommend co-stimulation with anti-CD28. With FluoroSpot Flex, it is possible to combine and build your own kit. For guidance look at our analyte combination table (above). Capture effects can be investigated by quantifying spot numbers in wells coated with single capture antibodies and compared to wells coated with a mixture of the capture antibodies. The compensatory effect of the anti-CD28 mAb may be assessed by comparing cells cultured with and without the anti-CD28 mAb.

Tutorial, Published December 28, 2023

Incubation strategies for success in ELISpot and FluoroSpot
We've made a handy table summarizing incubation times for different analytes to make your research planning for the ELISpot and FluoroSpot assay a little easier.
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