Abstract
The most challenging step in protein microarray fabrication is high-throughput production of proteins. Here we report two similar strategies to fabricate protein chips through capture onto a solid surface of the nascent polypeptides during translation of synthetic or in vitro–transcribed RNAs. Using these approaches, we efficiently fabricated both peptide and protein microarrays at relatively high density. We further demonstrated that such protein chips can be used to analyze protein activity.
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Acknowledgements
This research was supported by the National Institutes of Health (U54RR020839-01). We thank Jin Zhang for helpful discussions.
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S.C.T. performed all the experiments, H.Z. analyzed the data and both authors wrote the paper.
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The authors declare no competing financial interests.
Supplementary information
Supplementary Fig. 1
Protein chip fabrication. (PDF 1100 kb)
Supplementary Table 1
Oligos used in this study. (PDF 100 kb)
Supplementary Table 2
Effects of molar ratio of the BP oligo and synthetic peptide RNAs on the on-chip in vitro translation. (PDF 112 kb)
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Tao, SC., Zhu, H. Protein chip fabrication by capture of nascent polypeptides. Nat Biotechnol 24, 1253–1254 (2006). https://doi.org/10.1038/nbt1249
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DOI: https://doi.org/10.1038/nbt1249
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