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RISS 인기검색어
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- 저자 : Vandevyver, Caroline D. B. (검색결과 1 건)
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Ferná,ndez-Moreira, Vanesa,Song, Bo,Sivagnanam, Venkataragavalu,Chauvin, Anne-Sophie,Vandevyver, Caroline D. B.,Gijs, Martin,Hemmilä,, Ilkka,Lehr, Hans-Anton,Bü,nzli, Jean-Claude G. Royal Society of Chemistry 2010 The Analyst Vol.135 No.1
<P>The lanthanide binuclear helicate [Eu<SUB>2</SUB>(L<SUP>C2(CO<SUB>2</SUB>H)</SUP>)<SUB>3</SUB>] is coupled to avidin to yield a luminescent bioconjugate <B>EuB1</B> (<I>Q</I> = 9.3%, <I>τ</I>(<SUP>5</SUP>D<SUB>0</SUB>) = 2.17 ms). MALDI/TOF mass spectrometry confirms the covalent binding of the Eu chelate and UV-visible spectroscopy allows one to determine a luminophore/protein ratio equal to 3.2. Bio-affinity assays involving the recognition of a mucin-like protein expressed on human breast cancer MCF-7 cells by a biotinylated monoclonal antibody 5D10 to which <B>EuB1</B> is attached <I>via</I> avidin-biotin coupling demonstrate that (i) avidin activity is little affected by the coupling reaction and (ii) detection limits obtained by time-resolved (TR) luminescence with <B>EuB1</B> and a commercial Eu-avidin conjugate are one order of magnitude lower than those of an organic conjugate (FITC-streptavidin). In the second part of the paper, conditions for growing MCF-7 cells in 100–200 µm wide microchannels engraved in PDMS are established; we demonstrate that <B>EuB1</B> can be applied as effectively on this lab-on-a-chip device for the detection of tumour-associated antigens as on MCF-7 cells grown in normal culture vials. In order to exploit the versatility of the ligand used for self-assembling [Ln<SUB>2</SUB>(L<SUP>C2(CO<SUB>2</SUB>H)</SUP>)<SUB>3</SUB>] helicates, which sensitizes the luminescence of both Eu<SUP><SMALL>III</SMALL></SUP> and Tb<SUP><SMALL>III</SMALL></SUP> ions, a dual on-chip assay is proposed in which estrogen receptors (ERs) and human epidermal growth factor receptors (Her2/<I>neu</I>) can be simultaneously detected on human breast cancer tissue sections. The Ln helicates are coupled to two secondary antibodies: ERs are visualized by red-emitting <B>EuB4</B> using goat anti-mouse IgG and Her2/<I>neu</I> receptors by green-emitting <B>TbB5</B> using goat anti-rabbit IgG. The fact that the assay is more than 6 times faster and requires 5 times less reactants than conventional immunohistochemical assays provides essential advantages over conventional immunohistochemistry for future clinical biomarker detection.</P> <P>Graphic Abstract</P><P>Lanthanide luminescent bioprobes (LLBs) combined with microfluidics and lab-on-a-chip technology lead to fast dual assays of cancerous tissue biomarkers. <IMG SRC='http://pubs.rsc.org/services/images/RSCpubs.ePlatform.Service.FreeContent.ImageService.svc/ImageService/image/GA?id=b922124g'> </P>
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