Integrated barcode chips for rapid, multiplexed analysis of proteins in microliter quantities of blood

Abstract

As the tissue that contains the largest representation of the human proteome1, blood is the most important fluid for clinical diagnostics2,3,4. However, although changes of plasma protein profiles reflect physiological or pathological conditions associated with many human diseases, only a handful of plasma proteins are routinely used in clinical tests. Reasons for this include the intrinsic complexity of the plasma proteome1, the heterogeneity of human diseases and the rapid degradation of proteins in sampled blood5. We report an integrated microfluidic system, the integrated blood barcode chip that can sensitively sample a large panel of protein biomarkers over broad concentration ranges and within 10 min of sample collection. It enables on-chip blood separation and rapid measurement of a panel of plasma proteins from quantities of whole blood as small as those obtained by a finger prick. Our device holds potential for inexpensive, noninvasive and informative clinical diagnoses, particularly in point-of-care settings.

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Figure 1: Design of an integrated blood barcode chip (IBBC).
Figure 2: Measurement of human chorionic gonadotropin (hCG) in sera.
Figure 3: Multiplexed protein measurements of clinical patient sera.
Figure 4: IBBC for the rapid measurement of a panel of serum biomarkers from a finger prick of whole blood.

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Acknowledgements

The authors thank Larry Nagahara and Chris McLeland at the National Cancer Institute (NCI) for providing standard hCG serum samples and requesting the independent hCG measurement. We also thank Bruz Marzolf at the Institute for Systems Biology (Seattle) for printing DNA microarrays, and the UCLA nanolab for photomask fabrication. This work was funded by the National Cancer Institute grant no. 5U54 CA119347 (J.R.H., P.I.) and by the Institute for Collaborative Biotechnologies through grant DAAD19-03-D-0004 from the US Army Research Office.

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Authors

Contributions

R.F. developed and validated the DEAL barcode assay, measured cancer patient serum samples and analyzed all data. O.V. and B.K.H.Y. developed the blood separation chip. O.V., A.S. and L.Q. performed finger-prick blood test. R.F., O.V., A.S., H.A., G.A.K., C.-C.L. and J.G. participated in the synthesis and validation of reagents and the patterning of DNA barcode microarrays. L.H. and J.R.H. designed and directed the project.

Corresponding author

Correspondence to James R Heath.

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Competing interests

J.R.H. and L.H. are co-founders of Integrated Diagnostics, a company that has acquired the rights to license certain of the technologies described in this paper.

Supplementary information

Supplementary Text and Figures

Figures 1–13, Tables 1–3 (PDF 15420 kb)

41587_2008_BFnbt1507_MOESM2_ESM.wmv

This movie shows on-chip plasma separation by exploiting the Zweifach-Fung effect of highly polarized blood cell flow at branch points of small blood vessels. It is executed by flowing blood through the vertical primary channel that has high-resistance, centimeter-long channels branching off perpendicularly. As the resistance ratio is increased between the branches and the primary channel, a critical streamline moves closer to the primary channel wall adjoining the branch channels. Blood cells with a radius larger than the distance between this critical streamline and the primary channel wall are directed away from the high-resistance channels, and 15% of the plasma is skimmed into the high-resistance channels. The remaining whole blood is directed towards a waste outlet. (WMV 2099 kb)

Supplementary Video

This movie shows on-chip plasma separation by exploiting the Zweifach-Fung effect of highly polarized blood cell flow at branch points of small blood vessels. It is executed by flowing blood through the vertical primary channel that has high-resistance, centimeter-long channels branching off perpendicularly. As the resistance ratio is increased between the branches and the primary channel, a critical streamline moves closer to the primary channel wall adjoining the branch channels. Blood cells with a radius larger than the distance between this critical streamline and the primary channel wall are directed away from the high-resistance channels, and 15% of the plasma is skimmed into the high-resistance channels. The remaining whole blood is directed towards a waste outlet. (WMV 2099 kb)

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Fan, R., Vermesh, O., Srivastava, A. et al. Integrated barcode chips for rapid, multiplexed analysis of proteins in microliter quantities of blood. Nat Biotechnol 26, 1373–1378 (2008). https://doi.org/10.1038/nbt.1507

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