Antigen Binding and Site-Directed Labeling of Biosilica-Immobilized Fusion Proteins Expressed in Diatoms Public Deposited

http://ir.library.oregonstate.edu/concern/articles/r494vn13t

Supporting information is available online at:  http://pubs.acs.org/doi/abs/10.1021/acssynbio.5b00191

To the best of our knowledge, one or more authors of this paper were federal employees when contributing to this work. This is the publisher’s final pdf. The published article is copyrighted by American Chemical Society and can be found at:  http://pubs.acs.org/page/asbcd6/about.html

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  • The diatom Thalassiosira pseudonana was genetically modified to express biosilica-targeted fusion proteins comprising either enhanced green fluorescent protein (EGFP) or single chain antibodies engineered with a tetracysteine tagging sequence. Of interest were the site specific binding of (1) the fluorescent biarsenical probe AsCy3 and AsCy3e to the tetracysteine tagged fusion proteins and (2) high and low molecular mass antigens, the Bacillus anthracis surface layer protein EA1 or small molecule explosive trinitrotoluene (TNT), to biosilica-immobilized single chain antibodies. Analysis of biarsenical probe binding using fluorescence and structured illumination microscopy indicated differential colocalization with EGFP in nascent and mature biosilica, supporting the use of either EGFP or bound AsCy3 and AsCy3e in studying biosilica maturation. Large increases in the lifetime of a fluorescent analogue of TNT upon binding single chain antibodies provided a robust signal capable of discriminating binding to immobilized antibodies in the transformed frustule from nonspecific binding to the biosilica matrix. In conclusion, our results demonstrate an ability to engineer diatoms to create antibody-functionalized mesoporous silica able to selectively bind chemical and biological agents for the development of sensing platforms.
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  • Ford, N. R., Hecht, K. A., Hu, D., Orr, G., Xiong, Y., Squier, T. C., ... & Roesijadi, G. (2016). Antigen Binding and Site-Directed Labeling of Biosilica-Immobilized Fusion Proteins Expressed in Diatoms. ACS Synthetic Biology, 5(3), 193-199. doi:10.1021/acssynbio.5b00191
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  • description.provenance : Submitted by Patricia Black (patricia.black@oregonstate.edu) on 2016-04-21T14:59:49Z No. of bitstreams: 1 FordAntigenBindingSiteDirected.pdf: 1741155 bytes, checksum: 014559183f9050d7c4d8da94688a0878 (MD5)
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  • description.provenance : Approved for entry into archive by Patricia Black(patricia.black@oregonstate.edu) on 2016-04-21T15:00:12Z (GMT) No. of bitstreams: 1 FordAntigenBindingSiteDirected.pdf: 1741155 bytes, checksum: 014559183f9050d7c4d8da94688a0878 (MD5)

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