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Expanding a Portfolio of (FO‑) SPR Surface Chemistries with the Co(III)-NTA Oriented Immobilization of His6‑Tagged Bioreceptors for Applications in Complex Matrices
journal contribution
posted on 2020-04-08, 19:34 authored by Jia-Huan Qu, Sara Horta, Filip Delport, Machteld Sillen, Nick Geukens, Da-Wen Sun, Karen Vanhoorelbeke, Paul Declerck, Jeroen Lammertyn, Dragana SpasicCobalt-nitrilotriacetic
acid (Co(III)-NTA) chemistry is a recognized
approach for oriented patterning of His6-tagged bioreceptors.
We have applied the matching strategy for the first time on a surface
plasmon resonance (SPR) platform, namely, the commercialized fiber
optic (FO)-SPR. To accomplish this, His6-tagged bioreceptor
(scFv-33H1F7) and its target PAI-1 were used as a model system, after
scrutinizing the specificity of their interaction. When benchmarked
to traditional carboxyl-based self-assembled monolayers (SAM), NTA
allowed (1) more efficient FO-SPR surface coverage with bioreceptors
compared with the former and (2) realization of thus far difficult-to-attain
label-free bioassays on the FO-SPR platform in both buffer and 20-fold
diluted human plasma. Moreover, Co(III)-NTA surface proved to be compatible
with traditional gold nanoparticle-mediated signal amplification in
the buffer as well as in 10-fold diluted human plasma, thus expanding
the dynamic detection range to low ng/mL. Both types of bioassays
revealed that scFv-33H1F7 immobilized on the FO-SPR surface using
different concentrations (20, 10, or 5 μg/mL) had no impact
on the bioassay sensitivity, accuracy, or reproducibility despite
the lowest concentration effectively resulting in close to 20% fewer
bioreceptors. Collectively, these results highlight the importance
of Co(III)-NTA promoting the oriented patterning of bioreceptors on
the FO-SPR sensor surface for securing robust and sensitive bioassays
in complex matrices, both in label-free and labeled formats.
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Keywords
Complex Matrices Cobalt-nitrilotriacetic acidPAISAMplatformplasmagold nanoparticle-mediated signal amplificationFO-SPR sensor surfacebioreceptordifficult-to-attain label-free bioassaysNTAsurface plasmon resonanceFO-SPR surface coverageconcentrationscFv -33Hpatterningcarboxyl-based self-assembled monolayers
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