Lanthanide-Containing Polymer Microspheres by Multiple-Stage Dispersion Polymerization for Highly Multiplexed Bioassays
Identifieur interne : 000334 ( Ncbi/Merge ); précédent : 000333; suivant : 000335Lanthanide-Containing Polymer Microspheres by Multiple-Stage Dispersion Polymerization for Highly Multiplexed Bioassays
Auteurs : Ahmed I. Abdelrahman ; Sheng Dai ; Stuart C. Thickett ; Olga Ornatsky ; Dmitry Bandura ; Vladimir Baranov ; Mitchell A. WinnikSource :
- Journal of the American Chemical Society [ 0002-7863 ] ; 2009.
Abstract
We describe the synthesis and characterization of metal-encoded polystyrene microspheres by multiple-stage dispersion polymerization with diameters on the order of 2 µm and a very narrow size distribution. Different lanthanides were loaded into these microspheres through the addition of a mixture of LnCl3 salts and excess acrylic acid or acetoacetylethyl methacrylate (AAEM) dissolved in ethanol to the reaction after about 10% conversion of styrene, i.e., well after the particle nucleation stage was complete. Individual microspheres contain ca. 106 – 108 chelated lanthanide ions, of either a single element or a mixture of elements. These microspheres were characterized one-by-one utilizing a novel mass cytometer with an inductively coupled plasma (ICP) ionization source and time-of-flight (TOF) mass spectrometry detection. Microspheres containing a range of different metals at different levels of concentration were synthesized to meet the requirements of binary encoding and enumeration encoding protocols. With four different metals at five levels of concentration, we could achieve a variability of 624, and the strategy we report should allow one to obtain much larger variability. To demonstrate the usefulness of element-encoded beads for highly multiplexed immunoassays, we carried out a proof-of-principle model bioassay involving conjugation of mouse IgG to the surface of La and Tm containing particles, and its detection by an anti-mouse IgG bearing a metal-chelating polymer with Pr.
Url:
DOI: 10.1021/ja9052009
PubMed: 19807075
PubMed Central: 2801425
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<author><name sortKey="Baranov, Vladimir" sort="Baranov, Vladimir" uniqKey="Baranov V" first="Vladimir" last="Baranov">Vladimir Baranov</name>
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<front><div type="abstract" xml:lang="en"><p id="P1">We describe the synthesis and characterization of metal-encoded polystyrene microspheres by multiple-stage dispersion polymerization with diameters on the order of 2 µm and a very narrow size distribution. Different lanthanides were loaded into these microspheres through the addition of a mixture of LnCl<sub>3</sub>
salts and excess acrylic acid or acetoacetylethyl methacrylate (AAEM) dissolved in ethanol to the reaction after about 10% conversion of styrene, i.e., well after the particle nucleation stage was complete. Individual microspheres contain ca. 10<sup>6</sup>
– 10<sup>8</sup>
chelated lanthanide ions, of either a single element or a mixture of elements. These microspheres were characterized one-by-one utilizing a novel mass cytometer with an inductively coupled plasma (ICP) ionization source and time-of-flight (TOF) mass spectrometry detection. Microspheres containing a range of different metals at different levels of concentration were synthesized to meet the requirements of binary encoding and enumeration encoding protocols. With four different metals at five levels of concentration, we could achieve a variability of 624, and the strategy we report should allow one to obtain much larger variability. To demonstrate the usefulness of element-encoded beads for highly multiplexed immunoassays, we carried out a proof-of-principle model bioassay involving conjugation of mouse IgG to the surface of La and Tm containing particles, and its detection by an anti-mouse IgG bearing a metal-chelating polymer with Pr.</p>
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<title-group><article-title>Lanthanide-Containing Polymer Microspheres by Multiple-Stage Dispersion Polymerization for Highly Multiplexed Bioassays</article-title>
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<contrib-group><contrib contrib-type="author"><name><surname>Abdelrahman</surname>
<given-names>Ahmed I.</given-names>
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<contrib contrib-type="author"><name><surname>Dai</surname>
<given-names>Sheng</given-names>
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<contrib contrib-type="author"><name><surname>Thickett</surname>
<given-names>Stuart C.</given-names>
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<contrib contrib-type="author"><name><surname>Ornatsky</surname>
<given-names>Olga</given-names>
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<contrib contrib-type="author"><name><surname>Bandura</surname>
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<contrib contrib-type="author"><name><surname>Baranov</surname>
<given-names>Vladimir</given-names>
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<contrib contrib-type="author"><name><surname>Winnik</surname>
<given-names>Mitchell A.</given-names>
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<xref ref-type="corresp" rid="cor1">*</xref>
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<aff id="A1">Department of Chemistry, University of Toronto, 80 St George Street Toronto ON M5S3H6, Canada</aff>
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<author-notes><corresp id="cor1"><email>mwinnik@chem.utoronto.ca</email>
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<pub-date pub-type="nihms-submitted"><day>4</day>
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<pub-date pub-type="ppub"><day>28</day>
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<pub-date pub-type="pmc-release"><day>28</day>
<month>10</month>
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<volume>131</volume>
<issue>42</issue>
<fpage>15276</fpage>
<lpage>15283</lpage>
<abstract><p id="P1">We describe the synthesis and characterization of metal-encoded polystyrene microspheres by multiple-stage dispersion polymerization with diameters on the order of 2 µm and a very narrow size distribution. Different lanthanides were loaded into these microspheres through the addition of a mixture of LnCl<sub>3</sub>
salts and excess acrylic acid or acetoacetylethyl methacrylate (AAEM) dissolved in ethanol to the reaction after about 10% conversion of styrene, i.e., well after the particle nucleation stage was complete. Individual microspheres contain ca. 10<sup>6</sup>
– 10<sup>8</sup>
chelated lanthanide ions, of either a single element or a mixture of elements. These microspheres were characterized one-by-one utilizing a novel mass cytometer with an inductively coupled plasma (ICP) ionization source and time-of-flight (TOF) mass spectrometry detection. Microspheres containing a range of different metals at different levels of concentration were synthesized to meet the requirements of binary encoding and enumeration encoding protocols. With four different metals at five levels of concentration, we could achieve a variability of 624, and the strategy we report should allow one to obtain much larger variability. To demonstrate the usefulness of element-encoded beads for highly multiplexed immunoassays, we carried out a proof-of-principle model bioassay involving conjugation of mouse IgG to the surface of La and Tm containing particles, and its detection by an anti-mouse IgG bearing a metal-chelating polymer with Pr.</p>
</abstract>
<contract-num rid="GM1">R01 GM076127-03
||GM</contract-num>
<contract-sponsor id="GM1">National Institute of General Medical Sciences : NIGMS</contract-sponsor>
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<tree><noCountry><name sortKey="Abdelrahman, Ahmed I" sort="Abdelrahman, Ahmed I" uniqKey="Abdelrahman A" first="Ahmed I." last="Abdelrahman">Ahmed I. Abdelrahman</name>
<name sortKey="Bandura, Dmitry" sort="Bandura, Dmitry" uniqKey="Bandura D" first="Dmitry" last="Bandura">Dmitry Bandura</name>
<name sortKey="Baranov, Vladimir" sort="Baranov, Vladimir" uniqKey="Baranov V" first="Vladimir" last="Baranov">Vladimir Baranov</name>
<name sortKey="Dai, Sheng" sort="Dai, Sheng" uniqKey="Dai S" first="Sheng" last="Dai">Sheng Dai</name>
<name sortKey="Ornatsky, Olga" sort="Ornatsky, Olga" uniqKey="Ornatsky O" first="Olga" last="Ornatsky">Olga Ornatsky</name>
<name sortKey="Thickett, Stuart C" sort="Thickett, Stuart C" uniqKey="Thickett S" first="Stuart C." last="Thickett">Stuart C. Thickett</name>
<name sortKey="Winnik, Mitchell A" sort="Winnik, Mitchell A" uniqKey="Winnik M" first="Mitchell A." last="Winnik">Mitchell A. Winnik</name>
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