A myopic perspective on the future of protein diagnostics

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Abstract

Plasma proteome analyses of the future promise invaluable insights into states of health, not only by measuring proteins whose role it is to ensure blood homeostasis, but increasingly also as a window into the health of practically any tissue in the body via so-called leakage protein biomarkers. Realizing more of this vast potential will require progress along many lines. Here we discuss the main ones, such as optimal selection of target proteins, affinity reagents, immunoassay formats, samples, and applications, with a view from ongoing work in our laboratory.

Figures

  • Fig. 1. Commonly used immunoassay architectures. A. In reverse immunoassay, proteins in immobilized samples are recognized by labeled antibodies. B. In forward immunoassay, immobilized antibodies capture target molecules from labeled samples. C. In sandwich immunoassays, target proteins in a sample are capture by immobilized antibodies and detected by labeled antibodies added in solution phase. D. In proximity extension assays, target proteins in small aliquots of samples are bound by pairs of oligonucleotide-conjugated antibodies in solution phase. After an incubation the reactions are diluted to reduce chance proximity between reagents, and a DNA polymerase extends one or both oligonucleotides, templated by the other, to give rise to a reporter strand that can be quantified by realtime PCR. E. In solid phase proximity ligation assays, an immobilized antibody captures the target protein from a sample, followed by addition of pairs of oligonucleotide-conjugated antibodies. After washes, oligonucleotides in proximity are joined by ligation to give rise to amplifiable reporter DNA strands, quantifiable by realtime PCR.
  • Fig. 2. Dried blood spot sample collection. A. Only very small amounts of dried blood spot samples are required to interrogate by multiplex PEA. The smaller 1.2 mm diameter dot suffices for sensitive analysis of sets of 92 proteins. B. Results from a comparison of analyses of sets of 92 protein markers in 1 μl blood, or in 1.2mm diameter dots cut from dried blood spots on paper. Duplicate wet or dried blood samples (blue and red, respectively) were compared with each other. Results replotted from Ref. [36]

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CITATION STYLE

APA

Landegren, U., Al-Amin, R. A., & Björkesten, J. (2018, October 25). A myopic perspective on the future of protein diagnostics. New Biotechnology. Elsevier B.V. https://doi.org/10.1016/j.nbt.2018.01.002

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