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A sensitive and high-throughput assay to detect low-abundance proteins in serum

Nature Medicine volume 12, pages 473–477 (2006)Cite this article

Abstract

The ability to detect antigens immunologically is limited by the affinity of the antibodies and the amount of antigens. We have now succeeded in creating a modular, facile amplification system, termed fluorescent amplification catalyzed by T7 polymerase technique (FACTT). Such a system can detect protein targets specifically at subfemtomolar levels (∼0.08 fM). We describe here the detection of Her2 (also known as Neu) from rodent and human sera. FACTT is adaptable to high-throughput screening and automation and provides a practical method to enhance current ELISAs in medical practice.

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Figure 1: Illustration of FACTT.
Figure 2: Detection of streptavidin.
Figure 3: Detection of Her2.
Figure 4: Detection of serum Her2 in tumor-bearing mice.
Figure 5: Detection of Her2 from human sera.

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Acknowledgements

This work was supported by grants from the Abramson Family Cancer Research Institute to M.I.G. We thank X. (G.) Xu for the control sera from healthy volunteers and A. Massey for the preparation of the illustration for FACTT in Figure 1.

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Authors and Affiliations

  1. Pathology and Laboratory Medicine, School of Medicine, University of Pennsylvania, 3620 Hamilton Walk, Philadelphia, 19104, Pennsylvania, USA

    Hongtao Zhang, Xin Cheng, Mark Richter & Mark I Greene

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  1. Hongtao Zhang
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Correspondence to Mark I Greene.

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Zhang, H., Cheng, X., Richter, M. et al. A sensitive and high-throughput assay to detect low-abundance proteins in serum. Nat Med 12, 473–477 (2006). https://doi.org/10.1038/nm1378

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