ImmunoPheno: A Computational Framework for Data-Driven Design and Analysis of Immunophenotyping Experiments

Wu, L.; Nguyen, M. A.; Yang, Z.; Potluri, S.; Sivagnanam, S.; Kirchberger, N.; Joshi, A.; Ahn, K. J.; Tumulty, J. S.; Cruz Cabrera, E.; Romberg, N.; Tan, K.; Coussens, L. M.; Camara, P. G.

doi:10.64898/2026.02.01.703134

bioRxiv · 10.64898/2026.02.01.703134

ImmunoPheno: A Computational Framework for Data-Driven Design and Analysis of Immunophenotyping Experiments

Wu, L.·Nguyen, M. A.·Yang, Z.·Potluri, S.·Sivagnanam, S.·Kirchberger, N.·Joshi, A.·Ahn, K. J.·Tumulty, J. S.·Cruz Cabrera, E.·Romberg, N.·Tan, K.·Coussens, L. M.·Camara, P. G.

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Abstract

Immunophenotyping is fundamental to characterizing tissue cellular composition, pathogenic processes, and immune infiltration, yet its accuracy and reproducibility remain constrained by heuristic antibody panel design and manual gating. Here, we present ImmunoPheno, an open-source computational platform that repurposes large-scale single-cell proteo-transcriptomic data to guide immunophenotyping experimental design and analysis. ImmunoPheno integrates existing datasets to automate the design of optimal antibody panels, gating strategies, and cell identity annotation. We used ImmunoPheno to construct a harmonized reference (HICAR) comprising 390 monoclonal antibodies and 93 human immune cell populations. Leveraging this resource, we algorithmically designed minimal panels to isolate rare populations, such as MAIT cells and pDCs, which we validated experimentally. We further demonstrate accurate cell identity annotation across publicly available and newly generated cytometry datasets spanning diverse technologies, including spatial platforms like CODEX. ImmunoPheno complements expert curation and supports continual expansion, providing a scalable framework to enhance the accuracy, reproducibility, and resolution of immunophenotyping.

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Wu, L., Nguyen, M. A., Yang, Z., Potluri, S., Sivagnanam, S., Kirchberger, N., Joshi, A., Ahn, K. J., Tumulty, J. S., Cruz Cabrera, E., Romberg, N., Tan, K., Coussens, L. M., Camara, P. G.. 2026-02-03. ImmunoPheno: A Computational Framework for Data-Driven Design and Analysis of Immunophenotyping Experiments. https://doi.org/10.64898/2026.02.01.703134

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