We have two platforms for protein quantification from complex samples available. The first is a plated-based ELISA using Meso Scale Discovery’s electrochemiluminescence reader. The second is the AlphaLISA assay from Perkin Elmer which is faster but can not be done in multiplex. Both have their strengths and weaknesses so the project goals will determine which one is best. Both companies have local resources to help with new assay development.

Meso Scale: Assays provided by Meso Scale have 4-logs of linearity with detection limits in the low to sub-pico gram/ml range, depending on the assay. This assay can be multiplexed to as many as 10 assays/well on a 96-well Meso Scale plate. The ability to multiplex depends upon the expected antigen concentration range for each assay and potential cross-reactivity among Abs & Ags. The strengths are multiplexing of up to 42 samples in duplicate/plate, high sensitivity and a broad 4-log linear quantification range. The only drawbacks are the 6 hour total time to process a plate for the ELISA assays (mostly incubations and wash steps).

Perkin Elmer AlphaLISA:Premade assays are available from Perkin Elmer but primarily assays have to be developed by the end user. The assays are homogeneous, no wash steps and the time from sample to signal is much shorter for this assay. The assay can only be performed in singleplex but the shorter processing time compensates to a large degree. The assays have comparable dynamic range and are also quantitative.

Data Analysis: Data will be provided in an Excel workbook with the appropriate figures and post-run analyzed data. It is up to the investigator to complete data analysis in line with the experimental requirements.