Surface Functionalization Methods to Enhance Bioconjugation in Metal-Labeled Polystyrene Particles
Identifieur interne : 000488 ( Ncbi/Merge ); précédent : 000487; suivant : 000489Surface Functionalization Methods to Enhance Bioconjugation in Metal-Labeled Polystyrene Particles
Auteurs : Ahmed I. Abdelrahman [Canada] ; Stuart C. Thickett [Canada, Australie] ; Yi Liang [Canada] ; Olga Ornatsky [Canada] ; Vladimir Baranov [Canada] ; Mitchell A. Winnik [Canada]Source :
- Macromolecules [ 0024-9297 ] ; 2011.
Abstract
Lanthanide-encoded polystyrene particles synthesized by dispersion polymerization are excellent candidates for mass cytometry based immunoassays, however they have previously lacked the ability to conjugate biomolecules to the particle surface. We present here three approaches to post-functionalize these particles, enabling the covalent attachment of proteins. Our first approach used partially hydrolyzed poly(
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DOI: 10.1021/ma200582q
PubMed: 21799543
PubMed Central: 3143033
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<front><div type="abstract" xml:lang="en"><p id="P1">Lanthanide-encoded polystyrene particles synthesized by dispersion polymerization are excellent candidates for mass cytometry based immunoassays, however they have previously lacked the ability to conjugate biomolecules to the particle surface. We present here three approaches to post-functionalize these particles, enabling the covalent attachment of proteins. Our first approach used partially hydrolyzed poly(<italic>N</italic>
-vinylpyrrolidone) as a dispersion polymerization stabilizer to synthesize particles with high concentration of -COOH groups on the particle surface. In an alternative strategy to provide -COOH functionality to the lanthanide-encoded particles, we employed seeded emulsion polymerization to graft poly(methacrylic acid) (PMAA) chains onto the surface of these particles. However, these two approaches gave little to no improvement in the extent of bioconjugation. In our third approach, seeded emulsion polymerization was subsequently used as a method to grow a functional polymer shell (in this case, poly(glycidyl methacrylate) (PGMA)) onto the surface of these particles, which proved highly successful. The epoxide-rich PGMA shell permitted extensive surface bioconjugation of NeutrAvidin, as probed by an Lu-labeled biotin reporter (ca. 7 × 10<sup>5</sup>
binding events per particle with a very low amount of non-specific binding) and analyzed by mass cytometry. It was shown that coupling agents such as EDC were not needed, such was the reactivity of the particle surface. These particles were stable and the addition of a polymeric shell was shown did not affect the narrow lanthanide ion distribution within the particle interior as analyzed by mass cytometry. These particles represent the most promising candidates for the development of a highly multiplexed bioassay based on lanthanide-labeled particles to date.</p>
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<title-group><article-title>Surface Functionalization Methods to Enhance Bioconjugation in Metal-Labeled Polystyrene Particles</article-title>
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<contrib-group><contrib contrib-type="author"><name><surname>Abdelrahman</surname>
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<xref rid="A1" ref-type="aff">1</xref>
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Department of Chemistry, University of Toronto, 80 St George Street Toronto ON M5S3H6, Canada</aff>
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School of Chemistry F11, The University of Sydney, NSW 2006, Australia</aff>
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<abstract><p id="P1">Lanthanide-encoded polystyrene particles synthesized by dispersion polymerization are excellent candidates for mass cytometry based immunoassays, however they have previously lacked the ability to conjugate biomolecules to the particle surface. We present here three approaches to post-functionalize these particles, enabling the covalent attachment of proteins. Our first approach used partially hydrolyzed poly(<italic>N</italic>
-vinylpyrrolidone) as a dispersion polymerization stabilizer to synthesize particles with high concentration of -COOH groups on the particle surface. In an alternative strategy to provide -COOH functionality to the lanthanide-encoded particles, we employed seeded emulsion polymerization to graft poly(methacrylic acid) (PMAA) chains onto the surface of these particles. However, these two approaches gave little to no improvement in the extent of bioconjugation. In our third approach, seeded emulsion polymerization was subsequently used as a method to grow a functional polymer shell (in this case, poly(glycidyl methacrylate) (PGMA)) onto the surface of these particles, which proved highly successful. The epoxide-rich PGMA shell permitted extensive surface bioconjugation of NeutrAvidin, as probed by an Lu-labeled biotin reporter (ca. 7 × 10<sup>5</sup>
binding events per particle with a very low amount of non-specific binding) and analyzed by mass cytometry. It was shown that coupling agents such as EDC were not needed, such was the reactivity of the particle surface. These particles were stable and the addition of a polymeric shell was shown did not affect the narrow lanthanide ion distribution within the particle interior as analyzed by mass cytometry. These particles represent the most promising candidates for the development of a highly multiplexed bioassay based on lanthanide-labeled particles to date.</p>
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