Value of Cellular Proteomics: Completing the Functional Puzzle

Questions to ask during therapy development

There are many questions that arise when developing cellular, checkpoint, and bi-specific based immunotherapies, such as:

• What is differentiated about our therapy in this complex world of development?
• Which cells or source to use?
• Which bioprocessing method best suits our cell therapy?
• What is the actual correlative mechanism of response in our immunotherapy?

Each of these important questions rely on an understanding of whether your therapy is functioning the way you expect it to. The process to create and refine that cell or antibody-based therapy product begins in the early stages of lead generation. At this point, the question must be asked: do we have the most important pieces of the puzzle for understanding our impact on true immune cellular function?

Can you develop a therapy from the earliest stages to the clinic without knowing its true impact on immune proteomic function?

Without confirming how your immune therapy is functioning, how can you confidently determine a therapeutic is the optimal candidate? If you only know half of the puzzle, can you determine the effectiveness of your immune therapy?

A few challenges come to mind for establishing how your immune cells are functionally orchestrating response of various other immune cells via cytokines and chemokines as well as delivering the cytolytic payload to the tumor:

1. All immune cells, especially T cells, function heterogeneously, meaning that small, potent subsets of single cells might be missed in traditional bulk cytokine profiling measurements
2. RNA-seq correlates at 0.4 to the truly secreted cytokines per cell, meaning that measuring cytokines amongst many other targets at the RNA level is insufficient for defining true proteomic response
3. Finally, flow cytometry-based cytokine analysis estimates the true secretion of cytokines by blocking them within the cells, and is limited in multiplexing those blocked cytokines

How do we address these challenges for immunotherapy and complete the puzzle with new single-cell proteomic detection tools?

RNA-seq, histology, and flow cytometry have proven critical for tumor analyses and various mutational and location-based metrics. However, a true functional definition of immune fitness and function of each single cell is required to complete this puzzle to confirm how each immune cell is performing to discover robust correlations with response.

IsoPlexis is addressing this critical need and completing this puzzle as a new standard for functional cytokine immune profiling at the single-cell level. Additionally, IsoPlexis platforms have been able to correlate pre-infusion immune therapies with response, as published in Blood, as well as patient response to antibody based checkpoint therapeutics from the tissue and circulating T cells. With numerous case studies and over 60 publications and presentations, IsoPlexis has been proven as a new standard for this functional understanding of each cell within the development process.

IsoPlexis’ platform is the only source of this true functional profiling information, and overcomes the three challenges that exist today by:

1. Detecting the specific cytokines (over 30) that each single immune cell is secreting
2. Identifying the unique highly polyfunctional (secreting multiple cytokines simultaneously) subsets within a cell sample that are driving response, directly from the cytokine-based proteome of each cell
3. Characterizing true secretions per cell, thus defining correlative immune function, fitness, and potency of single cells

Using these metrics, IsoPlexis has generated data that has correlated with clinical outcomes in multiple studies, side by side, when legacy technologies such as flow cytometry and bulk ELISA have not.

Applying unique cellular functional data to development, highlighted as an emerging standard in Springer’s “Biomarkers for the Immunotherapy of Cancer”

In developing cell therapies, quickly determining how your cells are functioning and whether they are effective and potent means more accelerated and informed decisions, leading to better quality cell therapy, antibody based therapeutic products, and lead candidates. The IsoLight is the only system currently capable of this analysis and is becoming a standard tool in the drug development pipeline.

IsoPlexis’ IsoLight system was recently highlighted as a uniquely beneficial system for the standardization for Biomarkers for Immunotherapy of Cancer by Springer. This book of technical standards provides a deeper understanding of the scope of the biomarker methods to potentially improve the outcome from immunotherapies.

IsoPlexis was one of three immune phenotyping tools featured, and the only cellular functional analysis tool, selected as a biomarker in cell phenotyping. This is a testament to the data generated on the IsoPlexis platform as it continues to demonstrate the standardization of IsoPlexis’ systems for assessing correlative cellular fitness, determining optimal cell therapies in development, and providing advanced immune monitoring in checkpoint therapy, bispecifics, and more.

Single-cell functional proteomics is here and available now

In order to be certain that your cell therapy product and your antibody-based immunotherapies are functioning as predicted, measuring function at the single cell level is now a necessity (Figure). The only way to get this depth of functional information is with IsoPlexis’ platform.

WHAT’S MISSING?

WHERE ISOPLEXIS FITS

Traditionally, the most potent cells and the cytokines that determine function are missed through bulk averaging or other estimates

When you are not assessing the true function of your development products, questions regarding efficacy, fitness, potency, and correlations with cytokine mediated toxicities may go unanswered. For a complete picture of the drug development process, it is critical to functionally characterize your immune therapies at the single-cell level.

The proof is in the data: with true in vivo correlates, IsoPlexis’ technology is completing the puzzle in what is occurring within your therapies, so why wait? See how your cells are truly functioning. Let’s have a conversation.