%0 Journal Article
%A Herranz, Sonia
%A Marciello, Marzia
%A Olea, David
%A Hernández, Margarita
%A Domingo, Concepción
%A Vélez, Marisela
%A Gheber, Levi A.
%A Guisán, Jose M.
%A Moreno-Bondi, María Cruz
%D 2013
%T Dextran–Lipase
Conjugates as Tools for Low
Molecular Weight Ligand Immobilization in Microarray Development
%U https://acs.figshare.com/articles/journal_contribution/Dextran_Lipase_Conjugates_as_Tools_for_Low_Molecular_Weight_Ligand_Immobilization_in_Microarray_Development/2389894
%R 10.1021/ac400631t.s001
%2 https://ndownloader.figshare.com/files/4029583
%K platform
%K Microarray DevelopmentThe development
%K approach
%K waveguide
%K dextran
%K lipase
%K Modifying BTL 2
%K agarose macroporous beads
%K immobilizing
%K biosensor
%K immunosensor development
%K surface
%K conjugate
%K LR
%K IC
%K evanescent wave excitation
%K MCLR
%K Low Molecular Weight Ligand Immobilization
%X The development of
effective array biosensors relies heavily on
careful control of the density of surface-immobilized ligands on the
transducing platform. In this paper we describe the synthesis of new
dextran–lipase conjugates for use in immobilizing low molecular
weight haptens onto glass planar waveguides for immunosensor development.
The conjugates were synthesized by immobilizing bacterial thermoalkalophilic
lipases (Geobacillus thermocatenulatus lipase 2, BTL2) on agarose macroporous beads, followed by covalent
coupling to dextran networks of variable molecular weight (1500–40000).
The chimeras were immobilized via nonspecific hydrophobic interactions
onto glass planar waveguides modified with 1,1,1,3,3,3-hexamethyldisilazane
to obtain highly ordered and homogeneous molecular architectures as
confirmed by atomic force microscopy. Microcystin LR (MCLR) was covalently
bound to the dextran–BTL2 conjugates. The usefulness of this
approach in immunosensor development was demonstrated by determining
amounts of MCLR down to a few picograms per liter with an automated
array biosensor and evanescent wave excitation for fluorescence measurements
of attached DyLight649-labeled secondary antibody. Modifying BTL2
with dextrans of an increased molecular weight (>6000) provided
surfaces
with an increased loading capacity that was ascribed to the production
of three-dimensional surfaces by the effect of analyte binding deep
in the volume, leading to expanded dynamic ranges (0.09–136.56
ng L–1), lower limits of detection (0.007 ±
0.001 ng L–1), and lower IC50 values
(4.4 ± 0.7 ng L–1). These results confirm the
effectiveness of our approach for the development of high-performance
biosensing platforms.
%I ACS Publications