Abstract
Protein arrays are miniaturised and highly parallelised formats of interaction-based functional protein assays. Major bottlenecks in protein microarraying are the limited availability and high cost of purified, functional proteins for immobilisation and the limited stability of immobilised proteins in their functional state. In contrast, protein-coding DNA is readily available by PCR, and DNA arrays can be stored over prolonged times without deterioration. This chapter presents a method for the rapid and economical “printing” of replicate protein microarrays directly from a single DNA array template using cell-free protein synthesis, termed “DNA array to protein array,” DAPA. The procedure is a truly enabling technology, making customised protein microarrays affordable for laboratories with no access to routine microarray spotting. The experimental effort involved for the printing of a protein array from the template DNA array is comparable to the assembly of a Western blot.
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References
Hall D A, Ptacek J, Snyder M. Protein microarray technology. Mech Ageing Dev 2007;128:161–167.
Bertone P, Snyder M. Advances in functional protein microarray technology. FEBS J 2005;272:5400–5411.
He M, Taussig M J. Single step generation of protein arrays from DNA by cell-free expression and in situ immobilization (PISA method). Nucleic Acid Res 2001;29:e73.
Ramachandran N, Hainsworth E, Bhullar B, Eisenstein S, Rosen B, Lau A Y, Walter J C, LaBaer J. Self-assembling protein mircoarrays. Science 2004;305:86–90.
Angenendt P, Kreutzberger J, Glokler J, Hoheisel J D. Generation of high den-sity protein microarrays by cell-free in situ expression of unpurified PCR prod-ucts. Mol Cell Proteomics 2006;5:1658–1666.
He M, Taussig M J. DiscernArray™ technology: a cell-free method for the generation of protein arrays from PCR DNA. J Immunol Methods 2003;274:265–270.
He M, Stoevesandt O, Palmer E A, Khan F, Ericsson O, Taussig M J. Printing protein arrays from DNA arrays. Nat Methods 2008;5:175–177.
He M, Stoevesandt O, Taussig M J. In situ synthesis of protein arrays. Curr Opin Biotechnol 2008;19:4–9.
He M. Cell-free protein synthesis: applications in proteomics and biotech-nology. N Biotechnol 2008;Aug2025:126–132.
Khan F, He M, Taussig M J. A double-His tag with high affinity binding for protein immobilisation, purification, and detection on Ni-NTA surfaces. Anal Chem 2006;78:3072–3079.
Steinhauer C, Wingren C, Khan F, He M, Taussig M J, Borrebaeck C A. Improved affinity coupling for antibody microarrays: engineering of double-(His)6-tagged single framework recombinant antibody fragments. Proteomics 2006;6:4227–4234.
Acknowledgements
Research at the Babraham Institute is supported by Biotechnology and Biological Sciences Research Council (BBSRC), UK. The Protein Technology Group at Babraham Bioscience Technologies is a partner in the EC FP6 CA 026008 ProteomeBinders, and in.
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Stoevesandt, O., He, M., Taussig, M.J. (2011). Protein Microarrays Printed from DNA Microarrays. In: Khademhosseini, A., Suh, KY., Zourob, M. (eds) Biological Microarrays. Methods in Molecular Biology, vol 671. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-551-0_4
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DOI: https://doi.org/10.1007/978-1-59745-551-0_4
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