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Individually addressable parallel peptide synthesis on microchips

Nature Biotechnology volume 20, pages 922–926 (2002)Cite this article

Abstract

Miniaturized, spatially addressable microchips of peptides and peptidomimetics are powerful tools for high-throughput biomedical and pharmaceutical research and the advancement of proteomics. Here we report an efficient and flexible method for the parallel synthesis of peptides on individually addressable microchips, using digital photolithography and photogenerated acid in the deprotection step. We demonstrate that we are able to synthesize thousands of peptides in a 1 cm2 area on a microchip using 20 natural amino acids as well as synthetic amino acid analogs, with high stepwise yields and short reaction-cycle times. Epitope screening experiments using a p53 antibody (PAb240) produced clearly defined binding patterns. The peptidomimetic sequences on the microchip show specific antibody binding and provide insights into the molecular details responsible for specificity of epitope binding. Our approach requires just a conventional synthesizer and a computer-controllable optical module, thereby allowing potential development of peptide microchips for various pharmaceutical and proteomic applications in routine research laboratories.

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Figure 1: PGA deprotection.
Figure 2: Representative experiments for optimization of PGA deprotection.
Figure 3: PAb240 binding to the peptide and peptidomimetic microchip for synthesis validation and antibody-epitope analog discovery.
Figure 4: Results of positional scanning of the RHSVV epitope of p53 using PAb240 antibody binding.

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Acknowledgements

This research is supported by grants from the National Institutes of Health, National Cancer Institute, National Science Foundation, Texas ATP, the National Foundation for Cancer Research, and the R.A. Welch Foundation. We thank Ciro Nishiguchi for programming DIGI-SYN, Eric Leproust for sharing results on DNA chip synthesis and discussions, and Texas Instruments for the Digital Light Projector evaluation kits.

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Authors and Affiliations

  1. Department of Chemistry, University of Houston, Houston, 77004-5003, TX

    Jean Philippe Pellois & Xiaolian Gao

  2. Xeotron Co., 8275 El Rio, Suite 130, Houston, 77054, TX

    Xiaochuan Zhou & Tiecheng Zhou

  3. Department of Chemical Engineering, University of Michigan, Ann Arbor, 48109, MI

    Onnop Srivannavit & Erdogan Gulari

Authors
  1. Jean Philippe Pellois
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Correspondence to Xiaolian Gao.

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Pellois, J., Zhou, X., Srivannavit, O. et al. Individually addressable parallel peptide synthesis on microchips. Nat Biotechnol 20, 922–926 (2002). https://doi.org/10.1038/nbt723

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