As an add-on to our standard 51-plex panel, Enable Medicine offers custom workup for up to 3 additional biomarkers of your choosing. For each custom biomarker, you are welcome to suggest a particular clone for us to test. If you do not have a clone in mind, we will use our expertise to select one clone we think will work best. Then, the Enable lab will conjugate and test the selected clone in the CODEX assay.
For projects requiring more than 3 custom biomarkers, we will work with you to determine feasibility and specifics.
Enable Medicine provides a 2-step validation process for custom biomarkers. First, we perform an SDS-PAGE to confirm successful conjugation of the antibody to the CODEX barcode. In the rare case that conjugation fails, we will work with you to determine whether a second attempt is worthwhile.
Following successful conjugation of all custom markers, we perform one tissue test to evaluate custom marker performance in CODEX. This test will be run in normal tonsil tissue with our standard set of control markers (CD45, PanCk, Vimentin, aSMA, and CD31) unless otherwise specified. We will provide access to this test data in a QC study on our portal. QC study results are evaluated by our imaging scientists for appropriate co-staining, counter-staining, morphology, localization and abundance. Stains are compared to references when available, like those found on the human protein atlas. Once data is available, we will set a meeting to discuss the results and determine the best next steps.
The largest uncertainty in any custom project is lead time of reagents. For this reason, we do not begin workup until all custom components have been received. For example, in a project with 3 custom biomarkers, once we have received all three clones to be tested, and the necessary reagents for the conjugation reaction, we will be able to begin.
For up to 3 custom markers, we provide a two-week turnaround time to complete conjugation and share SDS-PAGE results. Following this, we require an additional four weeks to complete a test stain in CODEX. At this point, we will provide access to the test results in the Enable Medicine portal and reach out for next steps.
We are committed to providing consistent updates throughout the process of working together. You can expect updates as we receive reagents, confirm project kickoff, and have testing results available. In the event of any problems caused by unforeseen circumstances, we will work with you to determine our best course of action.
Overall, the expected timeline of custom biomarker workup and testing for 1 to 3 markers is 6 weeks from start to finish.
Determining antibody clones that work well with CODEX can be a lengthy process, dependent on many variables. We have the greatest success with clones that have been approved for use in IHC with FFPE tissue. The conjugation chemistry requires a purified antibody in a BSA-free formulation. We recommend PBS-only solutions. Solutions containing <0.1% sodium azide may be used. We do not recommend solutions containing glyerol. We recommend against purchasing clones that would require buffer exchange to meet these conditions. Many vendors can provide custom formulations upon request.
Even a clone that meets the strictest of these recommendations may not perform well in CODEX. Low expressing proteins can be particularly challenging, as CODEX technology does not offer signal amplification at this time.
If a clone tests unsuccessfully, we will work with you to decide whether to expand the scope of the workup project in order to test an additional clone. Once we have reached a final decision on all custom biomarkers, we are ready to receive study tissue and kick off the full study!
The cost of each custom biomarker project includes all necessary reagents and 2-step validation with shared results. The resulting volume of a single custom conjugation can be used for up to 8 staining experiments.