( strument were observed as shown in Figu...)Comparison of Performance between the AlphaQuest - HTS and the Fusion α Multilabel Reader for Detection of cAMP

Discussion and Conclusions

The AlphaQuest-HTS is a high throughput-screening instrument designed to measure exclusively AlphaScreen reagents in miniaturized formats. Although it is capable of measuring AlphaScreen samples in 96-, 384-well format for assay development purposes, it is not capable of measuring other assay methods. For this reason, the Fusion system was developed to offer versatility and AlphaScreen capability in one system. The versatility of the Fusion system provides the ability to create and optimize assays based on methods currently being used such as prompt fluorescence, absorbance, FP and luminescence. In addition, the Fusion can now be equipped with the AlphaScreen option, which creates the opportunity for assay developers to delve into this unique chemistry, while continuing to use current assay methods.

The significant difference between the Fusion α and the AlphaQuest-HTS for the detection of AlphaScreen assays is in the throughput capabilities as shown in Table 2. When the first AlphaScreen assay is ready to screen, the AlphaQuest-HTS incorporates two lasers and four PMTs to measure four samples simultaneously for screening purposes, and can measure almost 1000 samples/hour.

Graphical representations of the percent bound curves, similar IC₅₀ values, signal to noise, signal to background and ability to detect similar concentrations for an unknown sample confirm that the AlphaQuest-HTS and the Fusion α obtain the same results for a given assay. In addition, the regression analysis of raw data and percent bound values showed an extremely high correlation between the two instruments. This means that any assay that is developed on a Fusion α can be screened directly in the AlphaQuest-HTS without further development and optimization.

References