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<p>Existing high-plex protein measurement tools compromise on quantification, precision and cost efficiency. Here, to address this, we present nELISA, a platform that combines a DNA-mediated, bead-based sandwich immunoassay with advanced multicolor bead barcoding. Antibody pairs are preassembled on target-specific, barcoded beads, which ensures spatial separation between noncognate assays. Detection antibodies are tethered via flexible single-stranded DNA to enable efficient ternary sandwich formation. Detection is achieved through toehold-mediated strand displacement, where fluorescently labeled DNA oligos simultaneously untether and label detection antibodies. nELISA delivers sub-picogram-per-milliliter sensitivity across seven orders of magnitude. Using a 191-plex inflammation panel, we profiled cytokine responses in 7,392 peripheral blood mononuclear cell samples, generating ~1.4 million protein measurements and revealing over 440 robust cytokine responses, including previously unreported effects. nELISA thus provides a simple, scalable and cost-efficient solution for large-scale, high-fidelity phenotypic screening.</p>

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nELISA: a high-throughput, high-plex platform enables quantitative profiling of the inflammatory secretome

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年11月7日

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  • Milad Dagher,
  • Grant Ongo,
  • Nathaniel Robichaud,
  • Jinglin Kong,
  • Woojong Rho,
  • Ivan Teahulos,
  • Arya Tavakoli,
  • Samantha Bovaird,
  • Shahem Merjaneh,
  • Andrew Tan,
  • Kiran Edwardson,
  • Christelle Scheepers,
  • Andy Ng,
  • Andy Hajjar,
  • Baly Sow,
  • Michael Vrouvides,
  • Andy Lee,
  • Philippe DeCorwin-Martin,
  • Shafqat Rasool,
  • Jiamin Huang,
  • Timothy Erps,
  • Spencer Coffin,
  • Narges M. Rashidi,
  • Yu Han,
  • Srinivas Niranj Chandrasekaran,
  • Lisa Miller,
  • Maria Kost-Alimova,
  • Adam Skepner,
  • Shantanu Singh,
  • Anne E. Carpenter,
  • Jeffrey D. Munzar,
  • David Juncker

摘要

Existing high-plex protein measurement tools compromise on quantification, precision and cost efficiency. Here, to address this, we present nELISA, a platform that combines a DNA-mediated, bead-based sandwich immunoassay with advanced multicolor bead barcoding. Antibody pairs are preassembled on target-specific, barcoded beads, which ensures spatial separation between noncognate assays. Detection antibodies are tethered via flexible single-stranded DNA to enable efficient ternary sandwich formation. Detection is achieved through toehold-mediated strand displacement, where fluorescently labeled DNA oligos simultaneously untether and label detection antibodies. nELISA delivers sub-picogram-per-milliliter sensitivity across seven orders of magnitude. Using a 191-plex inflammation panel, we profiled cytokine responses in 7,392 peripheral blood mononuclear cell samples, generating ~1.4 million protein measurements and revealing over 440 robust cytokine responses, including previously unreported effects. nELISA thus provides a simple, scalable and cost-efficient solution for large-scale, high-fidelity phenotypic screening.