Jean-Claude Bunzli - Bioconjugated lanthanide luminescent helicates as multilabels for lab-on-a-chip detection of cancer biomarkers

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  Fernandez-Moreira,V., Song,B., Sivagnanam,V., Chauvin,A.S.,   Vandevyver,C.D.B., Gijs,M., Hemmila,I., Lehr,H.A., Bunzli,J.C.G.   Analyst 2010 135 (1) 42-52

  Abstract: The lanthanide binuclear helicate   [Eu(2)(L(C2(CO2H)))(3)] is coupled to avidin to yield a   luminescent bioconjugate EuB1 (Q 9.3%, tau((5)D(0)) = 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 EuB1 is attached via 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 EuB1 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 mu m   wide microchannels engraved in PDMS are established; we   demonstrate that EuB1 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(2)(L(C2(CO2H)))(3)] helicates, which   sensitizes the luminescence of both Eu(III) and Tb(III) ions, a   dual on-chip assay is proposed in which estrogen receptors (ERs)   and human epidermal growth factor receptors (Her2/neu) 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 EuB4 using goat anti-mouse IgG and   Her2/neu receptors by green-emitting TbB5 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

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