Abstract
To date, surface plasmon resonance (SPR) biosensors have been exploited in numerous different contexts while continuously pushing boundaries in terms of improved sensitivity, specificity, portability and reusability. The latter has attracted attention as a viable alternative to disposable biosensors, also offering prospects for rapid screening of biomolecules or biomolecular interactions. In this context here, we developed an approach to successfully regenerate a fiber-optic (FO)-SPR surface when utilizing cobalt (II)-nitrilotriacetic acid (NTA) surface chemistry. To achieve this, we tested multiple regeneration conditions that can disrupt the NTA chelate on a surface fully saturated with His6-tagged antibody fragments (scFv-33H1F7) over ten regeneration cycles. The best surface regeneration was obtained when combining 100 mM EDTA, 500 mM imidazole and 0.5% SDS at pH 8.0 for 1 min with shaking at 150 rpm followed by washing with 0.5 M NaOH for 3 min. The true versatility of the established approach was proven by regenerating the NTA surface for ten cycles with three other model system bioreceptors, different in their size and structure: His6-tagged SARS-CoV-2 spike fragment (receptor binding domain, RBD), a red fluorescent protein (RFP) and protein origami carrying 4 RFPs (Tet12SN-RRRR). Enabling the removal of His6-tagged bioreceptors from NTA surfaces in a fast and cost-effective manner can have broad applications, spanning from the development of biosensors and various biopharmaceutical analyses to the synthesis of novel biomaterials.
Highlights
Surface plasmon resonance (SPR) technology has been used for biosensing in various fields, spanning from the pharmaceutical analysis [1,2], medical and food diagnostics [3,4,5,6], to environmental monitoring [7]
Chemistry on the sensor probe surface, instead of istry [29]. This has opened up the potential for developing a surface regeneration approach and, asfrom such,our creating a reusable achieve this, we use originates
ethylenediaminetetraacetic acid (EDTA) has been included here in different buffer combinations since it is a stronger chelator than nitrilotriacetic acid (NTA), being essential to disrupt the chelate of NTA-Co(II)-His6 -tagged protein [36]
Summary
Surface plasmon resonance (SPR) technology has been used for biosensing in various fields, spanning from the pharmaceutical analysis [1,2], medical and food diagnostics [3,4,5,6], to environmental monitoring [7]. We have developed label-free sensitive compared to our previously developed immunoassays relying on EDC/NHS and sandwich immunoassays both in the buffer and complex matrices This has opened up the potential for developing a more surface fold diluted [29]. Co(II)-NTA chemistry on the FO sensor probe surface, instead of istry [29] This has opened up the potential for developing a surface regeneration approach and, asfrom such,our creating a reusable surface.
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