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Next-generation high-density self-assembling functional protein arrays

Nature Methods volume 5, pages 535–538 (2008)Cite this article

Abstract

We developed a high-density self-assembling protein microarray, based on the nucleic acid programmable protein array (NAPPA) concept, to display thousands of proteins that are produced and captured in situ from immobilized cDNA templates. We arrayed up to 1,000 unique human cDNAs and obtained high yields of protein expression and capture with minimal variation and good reproducibility. This method will enable various experimental approaches to study protein function in high throughput.

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Figure 1: Test array.
Figure 2: High-density array.
Figure 3: Protein interactions on high density arrays.

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Acknowledgements

We thank J. Williamson and M. Fernandez for their help with the robotics and D. Zhu and R. Boyce for developing the DNA normalization tool. This study was supported by the Early Detection Research Network (US National Cancer Institute grant 5U01CA117374-02) and the US National Institute of Allergy and Infectious Diseases (contract HHSN2332200400053C).

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Authors and Affiliations

  1. Harvard Institute of Proteomics, Harvard Medical School, 320 Charles Street, Cambridge, 02141, Massachusetts, USA

    Niroshan Ramachandran, Jacob V Raphael, Eugenie Hainsworth, Gokhan Demirkan, Manuel G Fuentes, Andreas Rolfs, Yanhui Hu & Joshua LaBaer

Authors
  1. Niroshan Ramachandran
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  2. Jacob V Raphael
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  3. Eugenie Hainsworth
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  4. Gokhan Demirkan
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  5. Manuel G Fuentes
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  6. Andreas Rolfs
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  7. Yanhui Hu
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  8. Joshua LaBaer
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Contributions

N.R. designed the experiment, processed slides and wrote the manuscript; J.V.R. and G.D. implemented automation, purified and printed DNA; E.H. analyzed array data; M.G.F. tested surface and printing chemistries; Y.H. performed informatics analysis of gene collection; A.R. cloned genes; and J.L. designed the experiment and wrote the manuscript.

Corresponding author

Correspondence to Joshua LaBaer.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–3, Supplementary Methods, Supplementary Protocol, Supplementary Tables 1–3 (PDF 846 kb)

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Ramachandran, N., Raphael, J., Hainsworth, E. et al. Next-generation high-density self-assembling functional protein arrays. Nat Methods 5, 535–538 (2008). https://doi.org/10.1038/nmeth.1210

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